Cervical cancer has high prevalence and mortality rates in worldwide female population. Persistent infection by high-risk Human Papillomavirus (hr-HPV) is the main cause of this cancer. However, many environmental, genetical, and epigenetical cofactors can modulate viral infection and cervical carcinogenesis. Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism is a genetic factor that has been associated with many pathologies, including cancer. Nevertheless, studies with cervical cancer presented controversial results, and varied according to ethnicity. Thus, the aim of this study was to determine association between MTHFR C677T polymorphism, Human Papillomavirus (HPV) infection and cervical cancer.n = 30), CIN II (n = 30), CIN III (n = 30), and Squamous Cervical Carcinoma (SCC) (n = 30). Control group was composed by 30 samples without lesion, presenting cervicitis. HPV detection was performed by conventional Polymerase Chain Reaction (PCR) with SPF primers set, and by real-time PCR specific for HPV 16 and hr-HPV. MTHFR C677T polymorphism was analyzed by PCR followed by Restriction Fragment Length Polymorphism (RFLP).A case-control study was performed with 150 histological cervical samples. Case group were divided in Cervical Intraepithelial Neoplasia (CIN) grade I (n = 109), CT 23.3% (n = 35) and TT 4.0% (n = 6). Polymorphic T allele frequency was 15.7%. No statistically significant association was observed between MTHFR C677T polymorphism and presence of pre-neoplastic or neoplastic cervical lesions. Similar frequencies of T allele was observed in control (23.3%) and cases (13.3%) groups (p = 0.174). In addition, there was no statistically significant association between MTHFR C677T polymorphism and viral infection, even considering hr-HPV or HPV 16 positivity.Frequency of MTHFR CC genotype was 72.7% (MTHFR C677T polymorphism was not associated with cervical cancer and HPV infection. Cervical cancer is the fourth most common cancer in women worldwide, with 529,000 new cases, and 275,000 deaths each year. Most cases occur in developing countries. On the other hand, lower incidence rates of this tumor occur in developed countries, where programs for prevention, screening and treatment are well established . In BrazPersistent infection with high-risk Human Papillomavirus (hr-HPV) is the main cause of cervical cancer, and 99.7% of cases are associated with the virus . HoweverMethylenetetrahydrofolate Reductase (MTHFR) is an enzyme that plays a central role in folate metabolism, important in cell replication and gene expression . C677T pMTHFR C677T polymorphism has been associated with development of cardiovascular diseases, defects in the neural tube formation, psychiatric disorders and various cancers . The preThus, the aim of this study was to determine association between MTHFR C677T polymorphism, HPV infection and cervical cancer in Brazilian women.Samples were collected between 2006 and 2011, from Brazilian women in Minas Gerais State, Brazil, a population with highly mixed ethnicities. Rating Histology of Richart was used to classify the samples . Tafuri n = 30), CIN II (n = 30), CIN III (n = 30), and SCC (n = 30). Thirty samples without cervical dysplasia was considered the control group . This assay is based on qualitative detection of hr-HPV , according to recommendations of manufacturer, and distinguish the presence of HPV 16.MTHFR C677T polymorphism was analyzed by PCR followed by Restriction Fragment Length Polymorphism (RFLP) Fig. , as descData were tabulated by Microsoft Office Excel®  and analyzed by Statistical Package for the Social Sciences® 17.0 .2 test following Monte-Carlo simulation were used to compare the groups.Descriptive statistics were performed to evaluate the frequency of MTHFR genotypes and HPV infection. Allelic frequency was calculated by Genepop software . FrequenTo evaluate the effect of age on the association between MTHFR C677T polymorphism and cervical carcinogenesis, samples were divided into two age groups using 30 years old as the cut off. After this age, oncogenic HPV infection is more persistent, and women are more likely to have pre-invasive lesion.Hardy-Weinberg Equilibrium (HWE) of MTHFR genotypes frequencies was calculated by Hardy-Weinberg equilibrium calculator including analysis for ascertainment bias .p values < 0.05 were considered as evidence of a significant association in all tests and 95% confidence interval was calculated when appropriate.Age range of women varied from 15 to 82 years, and mean and standard deviation of case and control groups were respectively 39.1 ± 13.5 and 40,5 ± 15.9 years.n = 109), 23.3% (n = 35) and 4.0% (n = 6) of CC, CT and TT, respectively. Frequency of polymorphic T allele was 15.7%. Distribution of genotypes was found under HWE .Genotypes frequencies of MTHFR C677T polymorphism were 72.7% (n = 145) of samples. Most of samples presented hr-HPV infection . Beside this, HPV 16 positivity was 28.0% (n = 42). However, no statistically significant difference was observed between distribution of MTHFR genotypes or alleles and HPV infection status  and alleles (p = 0.174), and presence of CIN or SCC (case group)   between distribution of genotypic and allelic frequencies and presence of cervical abnormalities according age group  directly transferred to deoxyuridine monophosphate (dUMP) in thymidylate synthesis; (b) oxidized to 10-formyltetrahydrofolate for de novo synthesis of purines; or (c) reduced to 5-methyltetrahydrofolate by MTHFR for methionine synthesis. Thus, reduction of enzymatic activity of MTHFR caused by C677T polymorphism may result in a lower synthesis rate of 5-methyltetrahydrofolate, leading to an increase availability of 5,10-methylenetetrahydrofolate for nucleotide production, which is essential for DNA synthesis and repair. In contrast, a lower proportion of 5-methyltetrahydrofolate is available for the methylation pathway, affecting genic expression , 21.However, although MTHFR C677T polymorphism may be linked to carcinogenesis by altering nucleotide synthesis, and DNA methylation pattern, in this study it was not possible to observe association between the presence of polymorphic genotype or allele, and cervical cancer. Other authors have also found no relationship between polymorphism in MTHFR gene and cervical carcinogenesis –31. HoweIn Brazil, only one study had been performed to evaluate the relationship between MTHFR C677T polymorphism and cervical cancer . The autNo association between HPV infection and C677T polymorphism in MTHFR gene were observed. We did not find other studies performed with Brazilian women, and analyzing association between MTHFR C677T polymorphism and viral infection. However, similar results were obtained with Indian population , 37. On Age is a cofactor that could also be considered. Development of cervical cancer begins with hr-HPV infection. If infection persists for 1 or 2 years, cervical abnormalities may occur. Only about 10% viral infection progress, leading to low-grade lesions. Approximately 2% of infected women have high-grade cervical lesions after years of persistent infection. High-grade abnormalities usually expand laterally around the transformation zone of cervical tissue, leading to invasive squamous cell carcinoma. Thus, there are more likely age groups for each stage of cervical cancer, with the highest prevalence of HPV infection occurring during adolescence and early adulthood, while the highest incidence of high-grade lesions is seen in women aged between 25 and 35 years. Carcinoma occurs predominantly between 45 and 60 years of age. Moreover, oncogenic HPV infection is more persistent in women aged 30 years or older, age group that women are more likely to have pre-invasive lesions .However, we did not observe significant difference in distribution of MTHFR genotype between cases and controls in both group ages evaluated (≤30 and > 30 years old). On the other hand, Sull and colleagues  concludeSome meta-analysis found that MTHFR C677T polymorphism is associated with cervical carcinogenesis differently according to ethnic group, acting as a risk factor for Asian, and as a protective factor for Caucasian , 14. HowBesides, cervical carcinogenesis is a multifactorial disease, although HPV infection is the main factor. Consequently, lack of information about other factors must be considered, like sexual behavior, multiparity, smoking and drinking habits, presence of other sexually transmitted diseases, and other genetic factors .Moreover, absence of data on serum folate of participants should be taken into account, since effect of MTHFR C677T polymorphism on HPV infection and development of cervical cancer can be modified by levels of folic acid , 42. MTHIn summary, no association between MTHFR C677T polymorphism, HPV infection and cervical intraepithelial neoplasia was found.


We investigated the association between bacterial microbiota in breast milk and the infant mouth. The influence of human papilloma virus (HPV) infection on infant oral microbiota was also assessed.Altogether 35 breast milk and 35 infant oral samples with known HPV status were selected from the Finnish Family HPV Study cohort. In total, there were 31 mother-infant pairs. The microbiota composition was characterized by 16S rRNA gene sequencing (V3-V4 region).Streptococcus, Staphylococcus, Unclassified Gemellaceae, Rothia, Veillonella, Haemophilus, Propionibacterium and Corynebacterium. HPV status was not associated with either microbiota richness or diversity in the infant mouth. However, the infant oral microbiota clustered in different groups according to HPV status. We detected higher abundance of Veillonella dispar (p = 0.048) at species level in HPV negative infant oral samples. We did not detect differences in the breast milk microbiota composition related to HPV infection due to only three HPV positive milk samples.HPV DNA was present in 8.6% (3/35) of the breast milk and 40% (14/35) of the infant oral samples. Eight shared genera between breast milk and infant oral were found; these included HPV infection is associated with distinct oral bacterial microbiota composition in infants. The direction of causality underlying the phenomenon remains unclear. The role of the bacterial microbiota in human health and disease has attracted considerable research interest during the last two decades. Much less is known about the interaction between viruses and the indigenous bacterial microbiota. Human papillomavirus (HPV) is a carcinogenic virus which has been causally associated with carcinomas in both the anogenital and the Streptococcaceae and Staphylococcaceae groups followed by Proteobacteria, Pseudomonadaceae, Bifidobacteriaceae[Breast milk is colonized with bacteria. Despite high intra- and interindividual variability, each mother harbors a unique and personalized microbial profile in her breast milk. The most predominant bacteria in human milk include teriaceae–7. The pteriaceae–11. Howeteriaceae et al. . BecauseHPV DNA was extracted by the high-salt method as described in Miller et al. . Singlethttp://cgenome.net/calypso/) was used with total sum normalization (TSS) for the statistical analysis. Linear discriminant analysis effect size (LEfSe) was used to detect unique biomarkers  score>3.0) in relative abundance of bacterial taxonomy. Inter and intraindividual variability was identified by pair-level analyses (paired t-test). We considered p-values ≤0.05 as statistically significant.Alpha diversity indices (Chao1 index: richness and Shannon index: diversity) and beta diversity using UNIFRAC (phylogenetic) and Bray Curtis distance (non-phylogenetic) were obtained. The Analysis of similarities (ANOSIM) test was used to test for statistically significant differences between the groups. Calypso software version 8.24 , Proteobacteria , Actinobacteria (5.2% vs 6.4%) and Bacteroides  (Streptococcaceae (50.7% vs 59.4%), Gemellaceae , Staphylococcaeae   . The pre 0.0078) . A core  0.0078) . This coStreptococcus, Veillonella and Rothia  was found in the two-month-old infant oral samples, whereas Lactobacillus, Bacteroides and Megasphaera  were higher in the infant oral samples collected at birth  are compared to those collected two months (n = 22) after birth . Furtherat birth .Micrococcaceae (p = 0.000051) and Streptococcaceae (p = 0.0051) were higher in samples taken two months after birth when compared to elevated level of Tissierellaceae (p = 0.033) in samples taken at birth. Furthermore, at species level Rothia mucilaginosa (p = 0.000043), Unclassified Streptococcus (p = 0.0051), Unclassified Weeksellaceae (p = 0.0095), Unclassified Haemophilus (p = 0.032) and Veillonella dispar (p = 0.034), Unclassified Anaerococcus (p = 0.030) and Unclassified Finegoldia (p = 0.052) levels were all higher at the age of two months compared to the samples taken at birth.At phylum level, the levels of Significant differences in the microbiota profile were found between breast milk and infant oral cavity using multivariate Redundant Discriminant Analysis (RDA) . When analyzing the microbiota data of the 31 mother- infant pairs we were not able to identify pairwise similarities or to connect mother to her infant by microbiota profile.There were only three HPV positive breast milk samples. No statistically significant differences in species richness  or diverProprionibacteriaceae (p = 0.095) was increased at family level, Propionibacterium (p = 0.095) at genus level and Propionibacterium acnes (p = 0.093) at species level in HPV positive infant oral samples. In the HPV negative infant oral samples, higher abundance of Veillonella (p = 0.025) at genus level and Veillonella dispar (p = 0.048) at species level was detected.  were significantly increased in the HPV positive group compared to the HPV negative infant oral sample group, whereas Veillonella  was higher in HPV negative infant oral samples  and Veillonella dispar (p = 0.052) levels were higher in the HPV negative infant oral samples collected two months after birth when compared to the HPV positive two months old infant’s oral samples.The impact of HPV infection on infant oral microbiota was investigated at two different time points albeit the sample size was relatively small. No differences in bacterial richness, diversity or composition were detected when comparing the infant oral HPV negative (n = 4) and the infant oral HPV positive (n = 8) samples collected at birth. However, we observed higher richness in the infant oral HPV negative samples collected two months after birth (n = 17) when comparing to the HPV positive infant oral samples at two months of age (n = 5). No differences in diversity were seen. Our results suggest for the first time that HPV infection is associated with alterations in the microbiota in the infant oral cavity. The changes are more pronounced at genus and family levels, but they do not influence species richness or diversity as a whole. Nonetheless, two distinct infant oral microbiota clusters dependent on HPV status were observed with RDA analysis. In contrast, we were not able to detect differences in breast milk microbiota composition associated with HPV infection, possibly because the total number of HPV positive breast milk samples was insufficient. Nevertheless, we still performed microbiota analyses, because the number of HPV positive milk samples 8.6%) were in line with previous studies.6% were .Streptococcus, Veillonella, Neisseria and Firmicutes[The infant oral microbiota observed in this study corresponds to previous studies, which indicate that the most abundant taxa in the infant oral microbiota belong to irmicutes,22. The irmicutes.While factors including prematurity, maternal intrapartum antibiotics, delivery mode and formula feeding all affect infant oral microbiota composition–32, the Veillonella, Unclassified Gemellaceae, Streptococcus, Staphylococcus, Rothia, Haemophilus and Cornyebacterium. These data are consistent with previous study by Biagi[Staphylococcus, Streptococcus and unclassified Gemellaceae shared between infant oral mucosa and breast milk microbiota. In our study, all of the infants received breast milk, though we are not aware, if there have been any additional food supply alongside breast feeding during the time when samples were collected. Because the main food supply has been from mother’s milk, we are confident that the results present an actual interaction between infant oral and breast milk microbiota.The breast milk microbiota is predominantly composed of bacteria which are thought to contribute to the infant’s newly developing gut microbiota and favorable microbiota prevents pathogens from colonizing the infant gut,5,34. ThPropionibacterium, Lysinibacillus and Ureaplasma were enriched in HPV positive infant oral mucosal samples when compared to HPV negative samples. Propionibacteriumin has been found in the gut microbiota of preterm infants[Ureaplasma has previously been linked to HPV positivity in sexually active men[Ureaplasma found in the cervix and placenta is connected to preterm birth without the presence of HPV[Ureaplasma has also been discovered in infant oral cavity collected 15 minutes after vaginal birth[Ureaplasma and Propionibacterium have previously been connected to preterm birth, our study had only total of three infants born slightly preterm and all of them were born after 35 weeks of gestation. Still it seems that HPV infection affects oral microbiota composition by favoring the growth of pathogens, which may have clinical significance later in life.The genera m infants. Oral Urctive men and Ureace of HPV. Recentlnal birth. DespiteVeillonella spp. was observed in HPV negative infant oral samples as compared to positive samples at two months of age. This finding is line with previous studies indicating that Veillonella is one of the most abundant bacteria in infant oral cavity [Veillonella has been associated with higher caries activity[An increase in the level of oral l cavity . Later i activity and poor activity.Proteobacteria, Streptococcaceae, Pseudomonadaceae, Bifidobacteriaceae and Staphylococcaceae[Lactobacillus have been reported to be higher in milk samples from mothers who delivered by caesarean section compared to vaginal delivery[Breastfeeding confers several significant health benefits to the infant. In addition, breast milk serves as one of the most important sources of microbes for the infant,34,41. Hcoccaceae–7, but tcoccaceae. Even de delivery. In our The Finnish family HPV study population and samples are exceptional. The study has been designed to evaluate HPV interactions within families, and consequently all of the samples have been collected within the same family. Nonetheless, our study has limitations which include the fact that only three breast milk samples were HPV DNA positive. The observed prevalence of 8.6% HPV positivity in breast milk is in line with previous studies. Still, Cyanobacteria, Chloroplasts and Rhizobiales or sequences that could not be classified to domain level were removed from the dataset.We have taken into consideration possible contamination and sequenced negative controls from PCR amplification and libraries. Furthermore, we aimed to verify our results by excluding singletons and OTUs with a relative frequency below 0.01 when analyzing of the data. In addition to that, sequences that were classified as In conclusion, present HPV infection in infant oral cavity is connected to detectable changes in microbial composition and it also seems to favor pathogens. Still, we do not know whether present HPV infection induces these changes in microbiota composition or whether the altered microbiota colonization predisposes to HPV infection. We were not able to find alterations in breast milk samples when taken into account present HPV infection. Finally, we conclude that infant oral and breast milk share similar features, as we found a core of eight shared bacteria.


Preferences of health professionals’ for human papilloma virus vaccines was measured by monetary value through willingness to pay (WTP) approach that could help policy makers set priorities among alternative cervical cancer prevention methods in poor countries. The objective of this study was to assess the female health professionals’ willingness to accept and pay, and associated factors for human papilloma virus vaccination at College of Medicine and Health Sciences, University of Gondar, Northwest Ethiopia.The majority (85.97%) of health professionals’ were willing to pay for Human papilloma virus vaccine. On the average, the respondents were willing to pay 231.34 ETB (US$8.50) per human papilloma virus vaccination service. Age, educational status, knowledge about cervical cancer and its risk factors and monthly income were identified as significant factors to WTP for human papilloma virus vaccination. Policy makers shall consider human papilloma virus vaccine to prevent cervical cancer maintain health of women’s and do more on rising awareness of individuals about cervical cancer and its risk factors. Cervical cancer is a chronic disease that arises due to the abnormal growth of cells from lower part of uterus which projects to the vagina . Among tEvery year, about 500,000 cervical cancer cases are diagnosed  and Sub-Loss of 1.5% of the world’s gross domestic product (GDP) associated with cancer due to mortality and incapacity without including direct medical cost of the treatment. In low-income countries burden of cancer was associated with sickness and death of mouth and throat cancer, cervix cancer, and breast cancer which were costing 1.3US$ billion, 1.3US$ billion and 1.1US$ billion respectively, in 2008 .Many countries in the world adopt human papilloma virus vaccine (HPV) as a strategy to overcome health and economic challenges which comes with cervical cancer .The government of Ethiopia had launched cervical cancer screening program in different site with the help of donors since 2013 and has a plan to introduce HPV in the country so as to prevent the incidence and negative impact of cervical cancer. To introduce HPV vaccine to Ethiopia, there is a need to study on willingness to accept and pay for HPV vaccine. Therefore this study was aimed to assess the acceptance rate of female health professionals for HPV vaccine and its benefit interim of money through willingness to pay approach.Besides, policy makers may know and look for optional strategies to tackle cervical cancer.Institutional based cross sectional study was conducted to investigate factors associated with willingness to accept and pay for HPV at the College of Medicine and Health Sciences, University of Gondar. The University of Gondar is a higher institution in Ethiopia located in northwest part of the country. The study was conducted from March 1st to April 30th, 2018.All female health professionals who were working at CMHS were the study population.The sample size of the study was calculated by using single population proportion formula for willingness to pay using the Epi Info™ 7 Stat Calc program with the assumption of 50% willingness to pay (p), 95% confidence level, 5% margin of error, and 10% response rate. The final sample size of the study was 423 female health professionals.The study participants were selected randomly by lottery method from the list of study population.An interviewer administered questionnaire was used to collect the data. Double-bounded dichotomous choice variant of the Contingent Valuation method and hypothetical scenario about HPV which was used to estimate the benefit of the program by eliciting the WTP of female health professionals. This method was used to estimate the benefit of the program in terms of monetary unit. The study participants were asked the maximum amount they were willing to pay for the service they got per visit.Willingness to pay is the maximum price the study participant will definitely be willing to buy per unit of a service.Contingent value method is an approach where the study participant was first asked whether they would be willing to pay 200 ETB per service to elicit the preference and then the question was repeated using a higher or lower bid value depending on the response to the first question until reaching to the final maximum amount the study participant will definitely buy.The data were entered into EpiData version3.1 software and was exported to STATA version14 software for analysis. Tobit model was used to estimate and identify determinants of willingness to pay. Tobit model is used to estimate outcome variables from censored data (skewed distribution of data). This model also predicts maximum amount of money that individuals were willing to pay for human papilloma virus vaccine. This model has an advantage over logistic discrete choice model in that it reveals the maximum amount of money the respondents were willing to pay.A total of 392 study participants, with a response rate of 92.7%, were included in the study. The mean age of the respondents was 28 years. Majority of respondents were Orthodox, Amhara, single, first degree holders and nurses Table . AverageIn this study 97.96% of respondents had heard about human papilloma virus vaccine. Schools were the main means of information about HPV vaccine for 60.68% respondents. Among the respondents 68.62% had excellent knowledge about cervical cancer danger signs and its risk factors (Table Most of respondents (85.97%) were willing to pay for human papilloma virus vaccination. Among participants 56.37% of them were willing to pay more than 200ETB and 31.63% were willing to pay more than 300ETB. The mean (± SD) amount of money health professionals willing to pay was 231.34ETB ± 132 per visit or 7.16 $USD.It’s calculated as follows: Currency exchange in March 2018 was 1$USD = 27.2ETB.The study showed that age, educational status, knowledge about cervical cancer and its risk factors and monthly income were factors statistically associated with willingness to pay for HPV vaccination in the multivariable analysis of Tobit model at a p-value ≤ 0.05 Table .Table 3FAs age increased by 1 year, respondents were willing to pay decreased by 2.5 Birr for human papilloma virus vaccination by holding other variables constant Table .First degree holders were willing to pay 63ETB less than those diploma holders for HPV vaccine by keeping other variables constant  . On the The average amount of money, the respondents willing to pay for HPV, was 231.34ETB (US$8.51) which was greater than the price used to elicit preference by 31ETB. The amount of money respondents were willing to pay for HPV was much higher than the amount of money respondents were willing to pay for injectable contraceptive per service and footwear for podoconiosis per year which were 11 Birr (US$0.65) and 64 Birr (US$ 3.28) , 14. TheThe age of respondents had negatively associated with willingness to pay. The negative association may be as age respondents’ increase, they may have less family income due to large family than younger age groups might be the possible explanations. This result was inconsistent with studies done on WTP for Pap test in Nigeria , willingThis study also showed that educational status was identified as significant variable to WTP for HPV. Possibly this might be as educational status increases, individuals might have more information and aware about their health issues. This report was consistent with studies done Addis Ababa , MalaysiThe knowledge of health professionals’ about cervical cancer and its risk factors had statistical association with willingness to pay. The possible explanation for this might be as individuals know more about cervical cancer and its risk factors, they become more willing to prevent cervical cancer. This finding was consistent with other studies conducted in Ethiopia , USA 2525 which The monthly income of the respondents’ had significant association WTP for human papilloma virus vaccination. This may be due to those having more income may had additional money to be allocated for promotion of their health in addition to basic needs. Studies conducted in Addis Ababa , MalaysiThis study indicated that majority of the health professionals were willing to pay for HPV Vaccination service. Age, educational status, knowledge about cervical cancer and its risk factors, and monthly income of the respondents were statistically associated with willingness to pay for HPV service.Policy makers shall consider human papilloma virus vaccine to prevent cervical cancer maintain health of women’s and do more on rising awareness of individuals about cervical cancer and its risk factors.The method used in this study is partial economic evaluation approach which it did not account other cervical cancer prevention methods. The study only includes educated female health professional might be the limitation of this study. Another limitation of this study was respondents’ bias that aims to reduce the cost of payment to the service and those have family history of cervical cancer need HPV vaccination might overestimate the cost.


Lactobacillus crispatus (multivariate OR (mOR) = 8.25, 95% CI 2.13~32.0) was predominant in the HPV-negative group. We observed that Eubacterium eligens , Gardnerella vaginalis , and Ureaplasma urealyticum  had the strongest associations with HPV clearance, and Lactobacillus johnsonii  with HPV persistence. Overall, greater diversity was observed in HPV-persistence than in HPV-negative women. Our findings suggest that the presence and prevalence of a specific cervical microbiome are factors involved in HPV dynamics.The present study aimed to identify the cervical microbes that are associated with HPV negativity, HPV clearance and HPV persistence and to assess the microbes’ longitudinal associations as related to HPV infection dynamics among Korean women. We enrolled 41 women with 107 samples, and classified them according to the HPV infection dynamics: HPV negativity , HPV clearance , and HPV persistence . Cervical swabs were collected at the baseline and six-month-interval follow-up visits. HPV positivity was determined by HPV DNA HC2 assay, and the microbiome was analyzed using 16SrRNA pyrosequencing, linear discriminant analysis effect size and multivariate logistic analysis. In the multivariate logistic analysis results,  The typical pattern of progression in HPV-mediated cervical cancer is as follows: (i) acquisition, (ii) persistence, (iii) progression to pre-cancer , and invasive cancer. There is evidence that most cases of HPV infection are transient, which is to say, likely to regress naturally3. Several cofactors such as smoking, high parity, long-term use of oral contraceptives6, hormone treatment and co-infection with sexually transmitted infection agents are relevant to progression of cervical cancer among HPV-infected women7. Associations between cervical microbes and HPV infection and CIN have been investigated9. Also, there have been reports on, for example, the frequency and determinants of acquisition and persistence of HPV infection among Danish soldiers10 and the viral and non-viral determinants of cervical HPV acquisition and clearance in Hawaiian women11. Recently too, an American study reported temporal changes of cervical microbes and HPV detection among Baltimore women12. Notwithstanding the many reports on cervical microbes and their HPV associations14, more knowledge on the epidemiology and longitudinal dynamics of cervical microbes is required before the nature of HPV progression is clearly understood. The present study aimed to identify the cervical microbes that are associated with HPV negativity, HPV clearance and HPV persistence and to assess the microbes’ longitudinal associations as related with HPV infection dynamics among Korean women.Cervical cancer is one of the leading causes of cancer death among women, and the most important causal agent in the development of cervical intraepithelial neoplasia (CIN) and cervical cancer is persistent infection with high-risk human papilloma virus (hrHPV)We enrolled 41 women and classified them into three HPV groups according to the HPV infection dynamics: HPV negativity , HPV clearance , and HPV persistence . The epidemiological and clinical information on the subjects is presented in Table Lactobacillus crispatus and Lactobacillus iners were the dominant species among the three HPV groups. A shift in the cervical microbiota was observed during the follow-up period (Supplementary Table Lactobacillus crispatus at the baseline (40.45%) and six-month-interval follow-up visits (35.6%). Compared with HPV-negative, HPV positive-women (HPV clearance + persistence group at baseline) had the highest abundance of Atopobium vaginae at the baseline and follow-up visits. Compared with HPV persistence, HPV clearance women showed increased percentage of Eubacterium eligens, Ureaplasma urealyticum at the baseline and follow-up visits. HPV persistence women showed high proportion of Lactobacillus johnsonii at the baseline and follow-up visits, compared with other groups (HPV negativity + clearance group).We sequenced 107 cervical samples (41 subjects) from a total of 617, 044 high-quality reads with an average of 18.97 operational taxonomic units (OTUs) per sample after quality filtering. The sequence reads were assigned to 14.5 OTUs in HPV negativity, 20.2 OTUs in HPV clearance, and 24.5 OTUs in HPV persistence. Eleven phyla — Firmicutes, Actinobacteria, Bacteroidetes, Fusobacteria, Proteobacteria, Tenericutes, Cyanobacteria, Verrucomicrobia, Planctomycetes, Acidobacteria, and Candidatus Saccharibacteria — were found, six of which  were dominant (>1%) among the groups. Gardnerella vaginalis  (P = 0.0403)  , an LDA score of at least 2, and a relative abundance greater than 0.1, both at the baseline and for total visits. The changes at the baseline are depicted in a cladogram Fig. . In the 03) Fig. , and the67) Fig. .Figure 1Lactobacillus crispatus, followed by Corynebacterium sundsvallense, Facklamia hominis, Fusobacteium naviforme, Antinobaculum schaali, and Helcococus ovis, with significant over-representation of Lactobacillus crispatus (P = 0.0016) , Eubacterium eligens (P = 0.0068) and Ureaplasma urealyticum (P = 0.0112)  Fig. . In the ted Fig. . The HPV12) Fig. .Figure 2We evaluated species richness (Chao 1 index), alpha-diversity (Shannon index) and beta-diversity of the cervical microbiome in baseline Fig.  and totaLactobacillus crispatus, Eubacterium eligen, Ureaplasma urealyticum, Gardnerella vaginalis and Lactobacillus johnsonii among the HPV groups. A multivariate logistic analysis (Table Lactobacillus crispatus (multivariate OR (mOR) = 8.25, 95% CI 2.13~32.0) was the most highly associated with HPV negativity. The multivariate OR of Eubacterium eligen for HPV clearance was higher  than for HPV negativity. Ureaplasma urealyticum  and Gardnerella vaginalis  were more highly associated with HPV clearance than with HPV negativity. Lactobacillus johnsonii  was associated more with HPV persistence than with HPV clearance.There were significant median differences in the relative abundances of is Table  showed tLactobacillus johnsonii, Haemophilus (genus) and Mycoplasmataceae (family) had the strongest association with HPV persistence, and that women with a cervical microbiome dominated by Eubacterium eligens, Gardnerella vaginalis and Ureaplasma urealyticum had the strongest associations with HPV clearance. Women with a high proportion of Lactobacillus crispatus were most likely to have HPV-negative infection. Higher bacterial diversity was observed in HPV-persistence women than in women showing HPV negativity.In the present study, we assessed the longitudinal associations of cervical microbes in HPV-negativity, clearance and persistence women. The main findings showed that the cervical microbiome differs significantly by HPV group. We found that women with high proportions of Lactobacillus johnsonii in HPV-persistent women.Based on these results of our two-year longitudinal study, we can suggest that bacterial dysbiosis is a factor associated with HPV dynamics (progression and regression). This study is, to the best of our knowledge, the first to examine the relationship between Haemophilus and Lactobacillus johnsonii was significantly associated  with HPV persistence among our Korean subjects. There is no previous study on Lactobacillus johnsonii in relation to cervical microbiomes, though it has been reported in saliva samples of HPV-positive and HPV-negative oropharyngeal cancer patients. A recent study by Guerrero-Preston et al. reported bacterial species Lactobacillus gasseri/johnsonii and Lactobacillus vaginalis in saliva of HPV-positive and HPV-negative oropharyngeal cancer patients15. However, the interaction between these microbes in saliva with HPV persistence and cancer risk was not discussed. The LEfSe analysis in the present study also confirmed enrichment of the genus Haemophilus and family Mycoplasmataceae in HPV-persistence women. The results for Mycoplasmataceae are in agreement with the finding of Abedamowo et al., that Mycoplasma hominis in cervical microbiota is significantly associated with hr-HPV infection16. It has also been reported that a few Mycoplasma species are efficient methylators and can promote cervical carcinogenesis through methylation of hr-HPV and cervical somatic cells16. One of the previous cervical microbiota studies (16-weeks samples) conducted by Brotman et al.12 demonstrated that high proportions of Atopobium spp. (belonging to CST IV-B) were associated with the slowest HPV clearance rate and that the persistence effect may be due to the high proportion of Atophobium vaginae that can play a role in the disruption of the epithelial barrier. The plausible molecular mechanisms of HPV persistence infection have been proposed: the viral life cycle and immune system evasion (avoidance of triggering of an immune response by the host)17, the tethering mechanism on host mitotic chromosomes 17, and E2 interaction with host receptors17. Then, 16 S rRNA gene sequencing revealed an increased proportion of L. johnsonii in HPV persistence and depletion of Lactobacillus crispatus relative to HPV-negative women; additionally, an alpha-diversity analysis provided further confirmation that HPV persistence in the total-visit samples showed increased species richness. Therefore, it can be suggested that changes in cervical microbiota with depletion in Lactobacillus crispatus and increased microbial diversity promote HPV infection and might be involved in HPV persistence18. The alpha-diversity results (Chao 1 index) showed increased diversity in HPV-persistence women relative to HPV-negative women in the total-visit samples. Therefore, high bacterial diversity in HPV persistence can represent an environment that increases the risk of progressing HPV persistence.Our multivariate logistic analysis confirmed that G. vaginalis, E. eligens and Ureaplasma urealyticum. The clearance effect might be due to the innate immune response (Toll-like receoptors-TLR) that is involved in a cascade of events promoting HPV clearance. TLR 3 and TLR9 play a role in regulating the pro-inflammatory cytokine and anti-viral environments of the lower female genital tract during viral and bacterial infection19; intense inflammatory response induced by G. vaginalis in viral clearance20, as well as viral proteins21 and other intrinsic host factors 22 responsible for HPV clearance. Another plausible explanation is that this clearance effect is due to an intense inflammatory response or that higher concentrations of inflammation markers (IP-10 and MIG chemokines) induced by bacteria in the cervical region assist in viral clearances23. One study conducted by Moscicki et al.20 reported that the HPV viral clearance effect might be due to the serendipity effect of Neisseria gonorrhe. One previous study24 reported the lowest clearance rate in women whose cervical microbiota were dominated by Atopobium and Gardnerella; therefore, further studies are needed to identify the mechanism or mechanisms whereby the composition of the cervical microbiota influence clearance.In the HPV-clearance women, multivariate logistic analysis and LEfSe analysis confirmed the significant enrichment of species Lactobacillus crispatus population remained stable at the baseline and follow-up visits, and Lactobacillus crispatus was significantly over-represented. Interestingly, in the LEfSe analysis results, we observed Corynebacterium sundsvallense, Facklamia hominis, Actinobaculum schaalii and Helcococcus ovis for the first time in the HPV-negative women. As reported earlier, in healthy women, hydrogen-peroxide producing Lactobacillus spp. are stable in maintaining the vaginal ecosystem25. Klebanoff et al. reported that Lactobacillus sp. increase TNF-α and IL-1α production, activate NF-κB in THP-1 cells and increase TNF-α production by human monocytes. This suggests that a higher concentration of Lactobacillus in the vagina can influence the physiology and host defenses therein25. In our previous study, we reported that Lactobacillus crispatus and Lactobacillus inners (Cluster III) were the dominant species in HPV-negative women and that high abundance of L. crispatus is associated with a low risk of CIN26.Among the HPV-negative women, the 29. Moreover, HC2 assay is technically well designed and can be easily controlled and performed by the laboratory technician. Whereas PCR-based method contains several steps that must be carefully optimized which make it more difficult to standardize PCR in laboratory condition.The strength of our present research is that the HPV samplings were longitudinal and that the differences between the cervical microbial compositions (species and genus level) among the HPV-negativity, clearance and persistence samples were identified by 16 S rRNA pyrosequencing. One of the most notable findings is that the bacterial communities in the cervical region of the HPV-persistence and clearance women were distinctive from those in the HPV-negative women, suggesting the presence of certain selective pressures contributing to the shift in the cervical microbiota. However, we also must acknowledge certain study limitations: (1) small sample sizes among the study groups, which could have biased the results; (2) an advanced-age study population (mean age: 44). So, the results should be interpreted with caution, and especially, it should be recognized that they might not apply to the general population; (3) another minor limitation of this study is HPV grouping by the liquid hybridization assay Hybrid Capture 2 (HC2) assays. It has been noted that HC2 assay shows some false positive results of 5%. Despite these limitations, HC2 assay has several advantages over other commercially available PCR-based target amplification methodL. crispatus species could be related to HPV persistence and also that the presence and prevalence of a specific cervical microbiome could be relevant to HPV dynamics.In conclusion, this study analyzed various cervical microbial taxa among different HPV groups, and found that increased bacterial diversity with reduced 30. The selected participants were interviewed using a structured questionnaire on their socio-demographic characteristics (Table 31. The samples were examined for the presence of one or more hrHPV types using the Hybrid Capture 2 (HC2) assay  according to the manufacturer’s instructions. The values obtained were recorded in relative light units (RLUs). All relative light units (RLU) measured on a luminometer were divided by the RLU of the respective positive control to provide a ratio. The sample was classified as a positive when the RLU/PC ratio was 1 pg/mL or greater. After baseline sampling, follow-up visits were scheduled at six-month intervals. The collected samples were stored at −80 °C for further analysis.We collected cervical samples from Normal and ASCUS patients (age between 18 and 65 years) who had participated in the Korean human papillomavirus (HPV) cohort study from 2006 to 2013. Detailed information on the inclusion/exclusion criteria for the HPV cohort is provided in our previous papernd visit, 20 at the 3rd visit, 10 at the 4th visit, and 1 at the final visit.To investigate the shifts of bacterial communities in the HPV states, we grouped the samples based on the following conditions: (1) HPV negative – persistently negative detection of hrHPV types throughout 24 months ; (2) HPV clearance – women who were initially hrHPV (baseline), and then regressed to negative during follow-up ; (3) HPV persistence – persistently positive detection of hrHPV types at the baseline and during follow-up   with signal amplification and chemiluminescence for 13 types of HR-HPV scored in RLU/PC. A positive result indicated a concentration of 1 pg/ml or higher than the RLU/cutoff ratio (RLU of the specimen/mean RLU of 2 positive controls).32. Beads recovered from emulsion PCR were deposited on a 454 Pico titer plate, and sequencing was executed using a Roche/454 GS-FLX plus. The sequencing run was performed by Macrogen Ltd. .DNA from cervical samples were isolated using the Fast DNA SPIN kit . The 16 S universal primers 27 F (5′ GAGTTTGATCMTGGCTCAG 3′) and 518 R (5′ WTTACCGCGGCTGC-TGG 3′) were used for amplification. A 20 ng aliquot of each sample was used for a 50 µl PCR reaction containing 10 × taq buffer, a dNTP mixture , 10 μm of the bar-coded fusion primers, and 2 U of taq Polymerase . The PCR conditions and pyrosequencing protocols are available elsewhere32. The primer sequences were cut down by employing pairwise sequence alignment, and sequences were gathered to correct for sequencing errors. Taxonomic identification was performed using the EzTaxon-e public database33 according to the highest pairwise similarity among the BLASTN search results. Possible chimera sequences were removed by the UCHIME algorithm34, and the diversity indices were calculated in Mothur after normalization of the read number in each sample. The potential biomarkers linked to HPV negative, HPV clearance and HPV persistence were analyzed by LDA effect size (LEfSe). Finally, the effect relevance was predicted by LDA35.The raw sequences for the samples were arranged using a unique barcode, and low-quality reads  were removed35. Beta diversity was calculated with principal coordinates analysis (PCoA) according to the Bray-Curtis distances. A permutational multivariate analysis of variance (PERMANOVA) was implemented to determine significance in distance. Diversity and the PERMANOVA results were analyzed using the R packages “vegan“36. The statistical analysis was performed with SAS 9.4, and R version 3.3.1 with ggplot2 packages was used for visualization37.The Chi-square test was used for testing of the categorical variables, and the Kruskal Wallis test was employed for comparison of the categorical continuous variables. When the number of expected frequencies was less than 5 and the number of cells was more than 25%, Fisher’s exact test was performed. To find a significant difference in alpha diversity, the Kruskal Wallis rank sum test was used to evaluate the differences in diversity among the three groups, followed by Dunn’s test of multiple comparisons. Multivariate logistic analysis was performed after adjusting for age, menopausal status, oral contraceptive use, and smoking habit. Risk estimates are presented as OR with 95% CI. LEfSe analysis was conducted to find significant differences among the relative abundance taxaThe Ethics Committee of the National Cancer Center approved this study, and all of the experiments were performed in accordance with the approved guidelines and regulations (IRB No. NCC2016-0147). Written informed consent was obtained from all of the study participants in accordance with good clinical practice.Supplementary Table S1


A simple, economical and metal-free approach to the synthesis of 2-substituted benzoxazoles and 2-substituted benzothiazoles from 2-aminophenols, 2-aminothiophenols and DMF derivatives, only using imidazolium chloride (50% mmol) as promoter without any other additive, was reported. Various 2-substituted benzoxazoles and 2-substituted benzothiazoles were thus prepared in moderate to excellent yields. Staphylococcus aureus sortase A Inhibitors oxazole (2a) oxazole (2b) oxazole (2c) oxazole (2d) oxazole (2e) oxazole (2f) oxazole (2g) oxazole (2h) oxazole (2i) thiazole (4a) thiazole (4b) thiazole (4c) thiazole (4d) thiazole (4e) thiazole (4f) thiazole (4g) thiazole (4h) thiazole (4i) thiazole (4j) [1H-NMR) δ 8.70 , 8.39 , 8.10 , 7.97 , 7.86 , 7.51 , 7.44–7.38 . 13C-NMR δ 169.35, 154.22, 151.37, 149.65, 137.05, 136.11, 126.30, 125.67, 125.29, 123.57, 122.02, 120.81. (See ole (4j) : The pro.81. See .3-Benzothiazol-2-yl-1-phenyl-propan-1-one (4k) [1H-NMR δ 8.02 , 7.95 , 7.83 , 7.57 , 7.50–7.40 , 7.34 , 3.66 , 3.57 . 13C-NMR δ 197.87, 170.59, 153.09, 136.50, 135.24, 133.36, 128.69, 128.14, 125.97, 124.85, 122.52, 121.54, 37.64, 28.29. (See one (4k) : The pro.29. See .


Anxiety affects over 260 million people worldwide. Benzodiazepines are a class of agents used in combination with other therapies for the management of anxiety. Lorazepam is a commonly prescribed benzodiazepine metabolized by uridine 5’-diphosphate-glucuronosyltransferases. Herein, we discuss recent findings regarding the pharmacogenetics of uridine 5’-diphosphate-glucuronosyltransferase 2B15 (UGT2B15), lorazepam, and its role in the treatment of anxiety. Anxiety is a common mental disorder affecting 264 million people worldwide, which includes generalized anxiety disorder (GAD), phobias, post-traumatic stress disorder, panic disorder, obsessive-compulsive disorder, and social anxiety disorder . In partUGT2B15*1/*1 (wild type) and UGT2B15*2/*2 (homozygous variant) genotypes in human subjects [UGT2B15*2/*2 group showed 40% to 50% lower systemic clearance of lorazepam during basal state compared to the UGT2B15*1/*1 group. Additionally, in the induced and inhibited states, the UGT2B15*2/*2 group showed lower absolute values of clearance. In their analysis, the researchers concluded that the UGT2B15 genotype accounted for 61% of the total variance of the systemic clearance of lorazepam, demonstrating a link between UGT2B15 genetic polymorphism and the pharmacokinetic profile of lorazepam. However, evaluation of clinically relevant parameters including sedation and psychomotor performance showed no significant difference in the two groups when subjects received lorazepam in the basal state.Uridine 5’-diphosphate-glucuronosyltransferases are responsible for the glucuronidation of lorazepam. Of interest, a recent study showed that genetic polymorphisms in the uridine 5’-diphosphate-glucuronosyltransferase 2B15 (UGT2B15) enzyme might affect lorazepam’s metabolism. Chung et al. looked at the differences in the pharmacokinetics and pharmacodynamics of lorazepam in subjects . Three mUGT2B15 polymorphisms and lorazepam pharmacokinetics [UGT2B15 genotype [UGT2B15*2/*2 group was much higher in the placebo group than the lorazepam group, whereas the lorazepam group showed greater anxiety reduction than the placebo group in UGT2B15*1/*1 and UGT2B15*1 /*2 (heterozygous variant) individuals. Interestingly enough, this relationship was the opposite in female patients, as females in the UGT2B15*2/*2 group showed greater anxiety reduction due to lorazepam premedication than placebo.Although Chung et al. found a clinically insignificant relationship between kinetics , researcUGT2B15 genotype may help clinicians decide when to prescribe lorazepam in certain patient populations in the future. Testing for the UGT2B15 genotype could potentially be a part of the larger clinical picture focusing on patient-centered care pending further evidence and cost considerations. Future research should focus on other types of anxiety exploring the effect of the UGT2B15 genotype on lorazepam.As shown in aforementioned studies, understanding the link between anxiety, lorazepam, and


IntroductionHuman papillomavirus (HPV) is the most common infection of the reproductive tract. The introduction of HPV vaccines by WHO aims to reduce the incidence of cervical cancer. Pakistan lacks an effective nationwide HPV vaccination program, thereby making HPV a major threat. In this study, we aimed to assess the knowledge of HPV vaccination in Pakistan and to determine the barriers against it.MethodsA cross-sectional study was conducted in Karachi, Pakistan, between April and May 2019. A convenience sampling technique was implemented using a self-administered questionnaire, which was filled by individuals aged 18-26. The questionnaire assessed the knowledge regarding HPV and also evaluated the attitude and acceptability amongst these individuals towards the vaccine. Data were analyzed using Statistical Package for Social Sciences (SPSS), version 20.0.ResultsThe majority of the participants belonged to the monthly household income range of 200,000 rupees and above, which was labeled as the high-income category . Out of the 18 people who were vaccinated, eight belonged to the aforementioned category. Of these 18, nearly two-thirds  had gotten vaccinated upon the recommendation of their doctor. A statistically significant difference was found only between those currently enrolled in universities and previously vaccinated for HPV (p=.047). Nearly half of the responders perceived the vaccine to be time-consuming  and overpriced .ConclusionThe vaccination rate is low in Karachi, Pakistan. Concentrated efforts involving the healthcare system should be made to raise awareness regarding HPV and its vaccine thereby reducing barriers to HPV prevention Human papillomavirus (HPV) is the most common infection of the reproductive tract . This viThe HPV vaccine has been recommended by WHO as the prime approach for the prevention of cervical cancer, and should ideally be administered prior to first sexual contact . In 2006Being a lower-middle-income country with a high burden of cervical cancer, Pakistan lacks an effective nationwide HPV screening and vaccination program. As a result, the Pakistani population may be unaware of there being such a vaccine. The potential of a vaccine to reduce disease burden relies on its acceptance and uptake by the community. Effective community health education and awareness-raising is a key component of any vaccination program .Furthermore, there have been very few studies related to HPV vaccination in developing countries as compared to developed countries worldwide ,11. Thusopenepi.com. Considering the population size of 1,000,000 with an anticipating frequency of 53%, a sample size of 383 was deduced at a 95% confidence interval. All individuals belonging to the age group of 18 to 26 years who could comprehend the questionnaire and of sound cognitive skills were included in the study, regardless of any other demographic limitation. A convenience sampling technique was implemented using a self-administered questionnaire. Certain methods were adopted to minimize bias, which included, but were not limited to, convenience sampling by each author covering different areas of Karachi and including participants from various socioeconomic backgrounds. Additionally, the questionnaire was peer-reviewed, and a pilot study was conducted.A cross-sectional study was conducted in Karachi, Pakistan, between April and May 2019. The sample size was calculated using Following an extensive literature review, a standardized questionnaire was designed and distributed among participants. The self-administered questionnaire provided careful instructions and a brief description of the motives behind the study. Informed consent was obtained from each respondent, with the choice of anonymity due to certain personal demographics being inquired. The questionnaire was designed to not only inculcate knowledge regarding HPV but also to evaluate the attitude and acceptability amongst these individuals towards the vaccine. It was divided into three main parts, which incorporated socio-demographics, knowledge, and attitude. The first part included details regarding age, sex, relationship status, household income, the field of study, and employment status. The second section addressed the pre-existing knowledge about HPV, its transmission, associated diseases, and certain personal attributes, including sexual activity, past vaccinations, and if female, Pap smear status. Thirdly, there was an inquiry regarding previously received HPV vaccination and the respondents' own opinions about its application, including availability, presumed side effects, and cost.Data were analyzed using Statistical Package for Social Sciences (SPSS), version 20.0. Categorical variables were expressed using frequencies and percentages. Continuous variables were presented as a mean and standard deviation. A chi-squared test with a 95% confidence interval was used to compare categorical variables.A total of 400 people participated in this study, out of which nearly three quarters were female  Table . The meaWhen asked about various aspects of HPV, more than 2/3rd of the participants responded with the correct answers, with 91% (n=364) of the participants correctly recognizing HPV’s ability to infect both males and females , the intention for a future HPV vaccine is seen in 62.0% of subjects (n=237). The resolve for a future HPV vaccination is higher in females . There is a statistically significant difference between the intention to achieve HPV immunization in the future and gender, with females more likely to get vaccinated (p<0.001). Moreover, from the population unvaccinated for HPV, 95.5% (n=365) claimed that they would consider getting vaccinated for HPV in the future if their doctors recommended it.Interestingly, the majority of the participants in our study had sufficient knowledge of HPV. This is in contrast to a similar study conducted by Zhuang et al. among female students studying in a tertiary institute in Singapore wherein upon administering a 14 point grading system to assess knowledge of HPV it was seen that the participants had insufficient knowledge with a median score of 7 . As expeAs predicted, females were more inclined to vaccinate themselves against HPV than men . The reaAlthough a notable number of participants were convinced regarding the safety, necessity, and efficacy of the HPV vaccine, thus eliminating the need to emphasize on vaccine safety, nearly half of them claimed that they did not have the time to get vaccinated and that the vaccine was expensive . A studyThere is a stigma associated with HPV due to its association with cervical cancer, leading to a lack of open dialogue surrounding reproductive health in low and middle-income countries, making it difficult and sometimes nearly impossible to overcome preventable issues concerning sexual health . There iLike all other studies, our study also has its limitations. Firstly, our sample size is not representative of the entire population as we used convenience sampling. Secondly, since this is a cross-sectional study, there is a risk of recall bias regarding certain questions, such as the sources of HPV-related information. Thirdly, there is also a risk of social desirability bias because this study was conducted on a sensitive topic, despite the participants having the option of filling out the questionnaire anonymously. Lastly, the sample size only consists of 400 students and can thus, reduce the generalizability of the results.According to our study, a significant portion of the population, especially females, were aware of the HPV vaccine and were inclined to get vaccinated as well. The men, on the other hand, were not as well-informed. However, the rate of vaccination uptake, for both genders, was not as high due to the exorbitant price, inaccessibility of the vaccine as well as the associated misconceptions that the use of the vaccine will encourage sexual promiscuity. Therefore, it is imperative that there is widespread awareness regarding the use of the vaccine targeted towards both genders. In addition, the vaccine should be readily available to all those who need it.


This diagnostic study compares the efficiency and effectiveness of 6 triage testing scenarios for screening women with high-risk human papillomavirus (HPV) for referral for colposcopy. What combination of human papillomavirus 16 (HPV-16) and HPV-18 genotyping with liquid-based cytological (LBC) testing is associated with the best performance for triaging women with HPV to detect cervical intraepithelial neoplasia ?This diagnostic study included 36 212 participants in the Forwarding Research for Improved Detection and Access for Cervical Cancer Screening and Triage study and found that a combined triage strategy of HPV-16/HPV-18 genotyping with reflex LBC was associated with an increase in the relative sensitivity to detect cervical intraepithelial neoplasia grade 2 or higher compared with LBC testing alone.The combination of HPV-16/HPV-18 genotyping with reflex LBC may offer a reliable strategy for discriminating women at greater risk of cervical cancer, avoiding unnecessary anxiety and diagnostic procedures in primary cervical cancer screening among women with high-risk HPV. Triage tests enhance the efficiency cervical cancer screening based on human papillomavirus (HPV), but the best approach for maximizing programmatic effectiveness is still uncertain, particularly in a real-world scenario.To compare the clinical performance of 6 triage strategies based on liquid-based cytology (LBC) and HPV-16 and HPV-18 genotyping individually or in combination as sequential triage tests to detect cervical intraepithelial neoplasia (CIN) grade 2 or higher among women with high-risk HPV.This diagnostic study of routine cervical cancer screening was conducted at 100 primary health centers in Tlaxcala, Mexico. Women aged 30 to 64 years were recruited from August 1, 2013, to February 24, 2016, as part of the Forwarding Research for Improved Detection and Access for Cervical Cancer Screening and Triage study. Six triage scenarios for referral to colposcopy were examined: (1) LBC testing that found atypical squamous cells of undetermined significance (ASC-US) or worse, (2) positive results in HPV-16 genotyping, (3) positive results in HPV-18 genotyping, (4) positive results in HPV-16/HPV-18 genotyping, (5) positive results in HPV-16 genotyping or, if genotyping results were negative, reflex LBC testing that found ASC-US or worse, and (6) positive results in HPV-16/HPV-18 genotyping or, if genotyping results were negative, reflex LBC testing that found ASC-US or worse. Data were analyzed from October 2017 to August 2018.Liquid-based cytological testing with simultaneous HPV-16 and HPV-18 genotyping. Women whose HPV genotyping results were positive for HPV-16 or HPV-18 or whose LBC results found ASC-US or worse and a random set of negative and normal results were referred to colposcopy with histologic analysis used for disease confirmation.Clinical performance of each test strategy for detection of CIN grade 2 or higher. Secondary outcomes included resource utilization of each triage scenario, measured by the number of tests performed, the referral rate for colposcopy, and the numbers of colposcopies per CIN grade 2 or higher detected.A total of 36 212 women  were screened, and 4051 women (11.2%) had high-risk HPV. Of these women, 1109 (24.6%) were found to have HPV-16, HPV-18, or ASC-US or worse. Further histologic testing detected CIN grade 2 or higher in 110 of 788 women (14.0%) who underwent follow-up colposcopy. Sensitivity and specificity for 3 main triage strategies were 42.9% and 74.0% for LBC; 58.3% and 54.4% for HPV-16/HPV-18 genotyping; and 86.6% and 34.0% for HPV-16/HPV-18 genotyping with reflex LBC. The referral rate to colposcopy was 29% for HPV-16/HPV-18 with reflex LBC, which was 2-fold higher than the referral rate of 12% for LBC.Triage of women with high-risk HPV with HPV-16/HPV-18 genotyping with reflex LBC was significantly associated with improvement in detection of CIN grade 2 or higher compared with LBC alone. The benefit of disease prevented may outweigh the cost of increasing requirements for colposcopy services in settings with limited adherence to follow-up after a positive screening result. A question that remains is the nature of the best combination of triage tests for cervical cancer screening using primary HPV testing.11 Recently, HPV-16/HPV-18 genotyping or cytological examination have been recommended to triage for immediate colposcopy in some national screening guidelines from high-income countries, offering greater sensitivity than cytological examination alone and better specificity than referring all women with HPV for colposcopy.14 Nevertheless, the suitability of these strategies in the routine health care practice in other resource-constrained settings has not been explored, to our knowledge. Country-based type-specific HPV prevalence and genotype distribution in cervical precancerous lesions,15 as well as the health care infrastructure relevant to LMICs, could influence the performance of these triage strategies.4A primary concern of testing for high-risk HPV, defined as an HPV infection with any of the 12 types of HPV that are known to cause cancer  to assess the performance and program components of alternatives for triaging women with HPV prior to integrating them into the national cervical screening program. In 2013, INSP launched the Forwarding Research for Improved Detection and Access for Cervical Cancer Screening and Triage (FRIDA) study, a demonstration project of HPV-based screening, that included the introduction of several tests for triaging to colposcopy with the goal of strengthening the current guidelines, which were launched in 2009. In this study, we aimed to compare the clinical test performance of the current triage tests available in Mexico, liquid-based cytology (LBC) testing and HPV-16 or HPV-18 genotyping, individually or in combination as sequential tests to detect cervical intraepithelial neoplasia (CIN) grade 2 or higher, among women with high-risk HPV participating in a routine cervical cancer screening program.STARD) reporting guideline.16The FRIDA study was approved by the institutional review boards of INSP, Tlaxcala State Ministry of Health, and the Mexican Federal Commission for the Protection Against Sanitary Risk. All participants provided verbal informed consent because the study involved no more than minimal risk and the same procedures of routine cervical cancer screening were used. This report adhered to the Standards for Reporting of Diagnostic Accuracy  LBC examination that found atypical squamous cells of undetermined significance (ASC-US) or worse, (2) positive results in HPV-16 genotyping, (3) positive results in HPV-18 genotyping, (4) positive results in HPV-16/HPV-18 genotyping, (5) positive results in HPV-16 genotyping or, if genotyping results were negative, reflex LBC testing that found ASC-US or worse, and (6) positive results in HPV-16/HPV-18 genotyping or, if genotyping results were negative, reflex LBC testing that found ASC-US or worse. Relative sensitivity and specificity of each triage strategy were estimated compared with LBC. The resource utilization for each triage strategy was a secondary outcome.Each participant underwent a pelvic exam to collect 2 cervical samples using 2 separate cervical brushes (Rovers). One brush was preserved in PreservCyt vial (Hologic) for HPV DNA and other molecular testing. The other brush was placed in a SurePath vial  for LBC testing.Vials for HPV DNA testing were transported to the INSP HPV laboratory in Cuernavaca, Mexico, and stored at 2°C to 8°C until testing could take place. Samples were tested for high-risk HPV using the cobas 4800 HPV test (Roche Molecular Diagnostics), a qualitative in vitro assay that identifies a pooled result for 12 high-risk HPV types, including HPV-31, HPV-33, HPV-35, HPV-39, HPV-45, HPV-51, HPV-52, HPV-56, HPV-58, HPV-59, HPV-66, and HPV-68, and individual results for HPV-16 and HPV-18.18 The threshold used to define abnormal cytologic results was ASC-US or worse.The vials used for LBC testing were stored at 2°C to 8°C upon arrival at the central cytology laboratory of Tlaxcala Health Services in Tlaxcala, Mexico. Samples were processed using the PrepStain System (TriPath Imaging) according to the manufacturer’s instructions. Slides were stained with the Papanicolaou method and interpreted according to the Bethesda 2001 criteria.Liquid-based cytology slides were reviewed by 2 independent cytotechnologists who were blinded to their counterpart’s result. If both cytotechnologists reported an abnormality, the worst result was established as the final diagnosis, otherwise a cytopathologist determined the final diagnosis. The cytopathologist also evaluated 5% of the slides with normal findings and all slides with ASC-US findings or worse, in compliance with the Mexican Cervical Cancer Screening program. Both the cytotechnologists and the cytopathologist were aware that all slides under evaluation were positive results for high-risk HPV but were blinded to HPV-16 and HPV-18 status.Women whose test results were positive in HPV-16 or HPV-18 genotyping or for whom LBC examination found ASC-US or worse were offered colposcopy, as well as randomly chosen women (29%) whose test results were negative for HPV-16/HPV-18 genotyping and whose LBC findings were normal, to correct for partial verification bias.19 Colposcopists were aware of the women’s triage test results and medical history. In addition, colposcopists collected a minimum of 4 biopsies (1 per quadrant of the squamocolumnar junction) and an endocervical sample in all women regardless of the colposcopic findings.All colposcopies were performed by 4 trained colposcopists in 2 colposcopy clinics, Tlaxcala’s General Hospital and Women’s Hospital. The colposcopic findings were reported according to the 2011 International Federation of Cervical Pathology and Colposcopy guidelines.20 If the diagnoses of the first 2 pathologists agreed, that was the final diagnosis; otherwise, biopsies were read in a blinded fashion by a third pathologist for end point adjudication. Clinical data collection was completed in October 2017.These biopsy specimens were processed at the central pathology laboratory in Tlaxcala General Hospital. Pathological diagnosis served as the criterion standard for the diagnosis of cervical precancer and cancer. Histological evaluation was performed by a panel of pathologists who reported the diagnosis according to Mexico’s Cervical Cancer Screening Program’s criteria.Descriptive statistics were used to summarize baseline characteristics. We evaluated the diagnostic performance of the triage strategies among participants with high-risk HPV in the FRIDA study using the baseline screening results.21 Relative sensitivities were also estimated from the GEE method with a log link function, using LBC alone as the comparator, which is the triage test used in the current screening program in Mexico.Two disease end points were assessed: (1) CIN grade 2 or higher, defined as women with CIN grade 2, CIN grade 3, or cancer confirmed via histologic analysis and (2) CIN grade 3 or higher, defined as CIN grade 3 or cancer confirmed via histologic analysis. For each triage scenario, we calculated sensitivity, specificity, positive predictive value, and negative predictive value, with 95% CIs using a generalized estimating equation (GEE) approach with an exchangeable working correlation structure to account for the within-woman correlation in triage results.24 To obtain verification bias adjusted (VBA) estimates of sensitivity and specificity as if all women with high-risk HPV had received the criterion standard, a weighted GEEs method was used.21 This method incorporates a weight representing the inverse probability of undergoing the criterion standard test, that is, the inverse of the sampling fraction of receiving the criterion standard test within women with high-risk HPV, either to allow VBA in women whose triage tests were negative and normal or to account for nonadherence among women with positive or abnormal triage test results referred to colposcopy.By design, partial verification bias occurred in our study, as only a small fraction of women with high-risk HPV who had a negative HPV-16 and HPV-18 genotyping results and normal LBC examination results were verified by histologic analysis, leading to missing data on disease outcome and biased estimates of diagnostic performance.P value of more than .05 was assumed to indicate that the sensitivity or specificity of the index algorithm was not significantly different from the comparator.25The McNemar test for discordant pairs was used to test the differences in sensitivity and specificity between paired triage results within CIN grade 2 or higher and CIN less than grade 2 groups. A McNemar 24 All analyses were 2-tailed, and a P value of less than .05 was considered significant. Statistical calculations were performed using Stata software version 14.1 (StataCorp). Data were analyzed from October 2017 to August 2018.To assess resource utilization of each triage scenario, we measured the number of tests performed, the referral rate for colposcopy, and the numbers of colposcopies per CIN grade 2 or higher detected. The numbers of colposcopies per CIN grade 2 or higher detected is an indicator of the diagnostic efficiency provided by each triage test, calculated as number of the histologically-confirmed CIN grade 2 or higher divided by the number of colposcopies performed.P for trend < .001). Most women were married or cohabiting (31 995 women [88.4%]), with age at sexual debut at least 18 years in 23 026 women (63.6%). The median (IQR) number of lifetime sexual partners was 1 (1-2) partner, with 25 408 women (70.2%) reporting having had only 1 lifetime partner. Most women had had at least 1 pregnancy (35 313 women [97.5%]), and nearly half had had 3 or 4 pregnancies (17 381 women [48.0%]). Only 101 women (0.3%) reported having received at least 1 HPV vaccine dose, and 14 977 women (41.4%) reported having undergone a Papanicolaou test within the previous 3 to 5 years  age of participants was 40 (35-47) years , with si 5 years .The STARD flow diagram is presented in the eFigure in the Of 3962 women with high-risk HPV and complete triage results, 1109 women (28%) had at least 1 positive or abnormal triage result, and 102 women (2.5%) had test results that were positive for HPV-16 or HPV-18 and ASC-US or worse cytologic abnormality. Among 1109 women with positive results for at least 1 of the triage tests, 788 women (71.1%) underwent colposcopy and had complete histological test results. The remaining 2853 women (70.4%) had negative results on HPV-16 and HPV-18 genotyping tests and normal findings on LBC tests, from which 366 of 818 women randomly chosen for VBA underwent colposcopy and had histological results. The characteristics of women in the control group for VBA and the rest of the women with negative or normal results on all triage tests are presented in eTable 2 in the P = .04) compared with LBC for detecting CIN grade 2 or higher. Conversely, LBC had higher specificity for CIN grade 2 or higher  than HPV-16/HPV-18 genotyping . The highest sensitivities for detection of CIN grade 2 or higher were found using the sequential algorithms of HPV-16 genotyping with reflex LBC  and HPV-16/HPV-18 genotyping with reflex LBC , although a comparison between these two sequential tests showed that HPV-16/HPV-18 genotyping with reflex LBC had statistically significantly higher sensitivity (McNemar P = .03). Lower specificities were observed in HPV-16 genotyping with reflex LBC  and HPV-16/HPV-18 genotyping with reflex LBC  compared with LBC as single test for triage. The To allow comparisons between different pairs of triage tests, a cross-tabulation is presented as eTable 1 in the Of all triage alternatives, the sequential triage tests were associated with the highest proportion of colposcopy referrals, including 939 referrals of 4048 women (23.2%) with the HPV-16 genotyping with reflex LBC triage scenario and 1109 referrals of 3962 women (28.0%) with the HPV-16/HPV-18 with reflex LBC triage scenario, whereas LBC alone resulted in 472 referrals of 3949 women (12.0%) . The str29 A sequential triage algorithm for screening women with high-risk HPV with HPV-16/HPV-18 genotyping as the first test may be a practical alternative for organizing HPV-based screening programs. Most validated high-risk HPV tests yield HPV-16/HPV-18 results as part of the same testing process and HPV-16/HPV-18 genotype testing is more reproducible than cytological analysis. Moreover, it has been demonstrated that presence of HPV-16 is a major factor for risk of CIN grade 3 or higher.31 A 2016 cohort study31 found that HPV-16 and HPV-18 are associated with the greatest 3-year risk of CIN grade 3 or higher.One of the most pressing issues in cervical cancer screening is finding adequate triage methods for patients with high-risk HPV. Thus, we assessed the clinical performance of LBC, HPV-16 genotyping, and HPV-18 genotyping for detection CIN grade 2 or higher using 6 different algorithms. The use of sequential testing based on HPV-16/HPV-18 genotyping followed by reflex LBC offered the highest sensitivity but the lowest specificity for detection of CIN grade 2 or higher compared with LBC alone, findings that corroborate those of previous cross-sectional screening trials.26 , our performance estimates for triage based on HPV genotyping were very similar. According to a study by Mayrand et al,32 sensitivity of HPV testing is less likely to be affected by identification of more incipient lesions through a multibiopsy approach. In contrast, we observed a more striking difference for the LBC triage sensitivity, with 42.9% in our study vs 52.6% obtained in the ATHENA HPV study.26 Assuming that colposcopic findings are what influence a colposcopist to target cervical areas for biopsy, visible and mostly larger lesions would likely be associated with the higher sensitivity estimates for LBC found in earlier studies. Additionally, although we conducted a prestudy training for interpreting LBC slides, subjectivity plays an important role in the performance of the tests, as noted by a subanalysis of the ATHENA HPV study26 by Castle et al.33 Problems inherent with misclassification of cytologic diagnosis, either by clinician factors, such as the variability of technical training and experience of our cytotechnologists, or by the cytological scoring itself, may also contribute to these differences.It is important to highlight that although a different biopsy protocol was applied in the Addressing the Need for Advanced HPV Diagnostics (ATHENA) HPV study34Our analysis found that decreasing the screening start age to offer HPV-based screening rather than LBC from 35 years to 30 years may be associated with an increase in the detection of precancerous lesions, as this age group contributed to one-third of the total CIN grade 2 or higher observed. Although this finding suggests the use of a more sensitive screening method, a key concern is the need to be less aggressive in CIN detection and management considering the potential for adverse reproductive outcomes associated with treatment of precancerous lesions in young women.An important implication of our findings for the application of these triage approaches in other settings is in the utilization of diagnostic resources. The HPV-16/HPV-18 genotyping with reflex LBC strategy was associated with an increase in the detection of CIN grade 2 or higher but also increased 2-fold the number of colposcopy referrals compared to LBC alone. Although the economic costs for follow-up, colposcopy, and treatment of false positives may seem sufficient reason to accept the failure to detect some CIN grade 2 or higher, especially if a significant proportion of lesions will spontaneously regress, these may be offset by the economic and social cost savings of preventing the occurrence of some cancers.35 we believe that treating a woman at high risk of cervical cancer will produce a better outcome for women’s overall health and for public health in general in the long term. Moreover, screening algorithms with low sensitivity require frequent repetitions of screening, which pose logistical barriers in LMICs that have barriers to adherence to follow-up recommendations as well as inequity in access to services.37Since women in resource-poor settings have a low probability of getting in contact with prevention services,38 Indeed, as vaccinated cohorts enter into screening, such biomarkers will be imperative to adjust clinical practices in accordance with the change in HPV patterns and the increasing number of less relevant lesions.39 For now, while HPV vaccination programs leave adult women unprotected, strategies based on HPV-16 and HPV-18 combined genotyping or LBC may offer an acceptable performance for triaging women with high-risk HPV.A more accurate detection of cancer progression through novel specific biomarkers based on DNA methylation or cell cycle dynamics might be influential in decisions to refer to or defer treatment.This study is the first large study evaluating triage alternatives in real-world LMIC conditions, to our knowledge. Although training was conducted prior to the beginning of our study, the study procedures were performed within a routine screening program, providing information about the triage performance in real-world environments. Our findings may thus be generalizable to populations with similar characteristics. The design of the FRIDA study permitted an efficient and unbiased comparison of triage tests and their algorithms. One of the main strengths was that our study included a systematic collection of biopsies, reducing the potential bias of differential disease misclassification associated with the colposcopic impression. It is worth noting that disease ascertainment was by pathology panel review, which was led by an experienced pathologist and attained a consensus diagnosis in all cases.40 This measurement error could be higher in the group of women whose results were negative in HPV-16/HPV-18 genotyping and normal in LBC who were used for VBA, considering that CIN grade 2 lesions caused by high-risk HPV types other than HPV-16 or HPV-18 are more likely to regress.42 A 2016 study31 indicated that the 3-year risk of CIN grade 3 or higher ranged from 1.2% to 4.0% for patients with high-risk HPV who did not have HPV-16 or HPV-18 and had normal cytological findings. This can lead to an underestimation of the true sensitivity and specificity of the analysis if using VBA. Although VBA estimates were reported to ensure comparability with other screening trials, our results should be interpreted with caution to prevent dissemination of misleading data about the potential benefit of a particular triage strategy.Our study had some important limitations. The FRIDA study was not powered to detect differential test performance characteristics by subgroups. We could not ascertain if some CIN grade 2 or CIN grade 3 that were detected at baseline may have been transient lesions, resulting in outcome misclassification.This diagnostic study found that HPV-16/HPV-18 genotyping with reflex LBC for triaging women with high-risk HPV is an appropriate method to achieve the goal of preventing cervical cancer in settings where access to appropriate follow-up is challenging, assuming that the costs and burden of overtreatment are offset by the individual and social benefits of preventing cervical cancer. Perceptions and preferences of women and health care professionals, as well as the resources required, are key factors that should be addressed to ensure effective screening programs.


The multiple visits required for an effective Pap smear screening program is difficult to replicate in many developing countries. This precludes early diagnosis and care for patients with cervical cancer and contributes to its high mortality in these countries. HPV screening has higher specificity and high negative predictive value and has the advantage that materials can be self-collected, which permits the screening of women who for various cultural and religious reasons would be reluctant to come to the clinic to expose themselves for screening. The aim of the study was to assess the degree of agreement between self sampling for HPV DNA with samples collected by a health provider.Each respondent selected from women presenting for cervical cancer screening underwent both self- and provider sampling for HPV DNA testing using Hybribio GenoArray.Of the 194 women screened, 12 (6.2%) and 19 (9.8%) had HPV on self sampling and provider col-lected samples respectively. The commonest HPV type seen using both techniques was HPV 58 (2.6%). Multiple HPV genotypes were seen in 1 (0.5%) and 5 cases (2.6%) of provider and self-collected samples respectively. The high risk-HPV detection rate was 7.2% when self sampled and 6.8% when sampled by the provider. There was moderate correlation between both sampling techniques .Our study shows moderate correlation between both sampling techniques. Larger multicentre studies will be needed to provide results generalisable to the Nigerian population. Keywords: Pap smear, HPV screening, cervical cancer, sample collection, self-sampling, provider collected, PCR, HPV DNA, Ile-Ife Nigeria. Cancer of the cervix is the third leading cause of cancer deaths in women worldwide following breast and colorectal cancers. Over the past 30 years however the incidence and mortality rates have declined in developed countries. This is largely due to effective and efficient cervical cancer screening programs using pap smears and HPV DNA testing . TestingStudy population: The study was conducted at the Gynaecological Oncology Clinic at Obafemi Awolowo University Teaching Hospital Complex (OAUTHC) Ile-Ife, a community based clinic for screening of cervical cancer as well as other diseases of the female genital tract. Informed consent was obtained from the women and ethical clearance was obtained from the Ethical Committee of OAUTHC.Sample collection: Systematic random sampling technique was used to stratify patients into two groups. Respondents in group A underwent provider sampling before self sampling, while respondents in group B had self sampling before undergoing provider sampling. All respondents underwent both sampling procedures. Both self collected samples and provider collected samples were obtained using cytobrush (cervexR) cervical cell sampler. Provider collected specimens were obtained by sampling the squamocolumnar junction using a 360° rotator movement of the cytobrush after the introduction of a speculum. The respondents were taught how to perform the procedure for sampling the upper vagina according to the instructions on the sample collection kit. Both specimens were dropped in the Hybribio HPV DNA collection kit provided for HPV DNA test.HPV genotyping: The samples were frozen at -20°C and sent to the College of Medicine University of Lagos for HPV typing which was done using HPV GenoArray test kits  for 21 HPV genotypes which included high risk types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68, low risk types 6, 11, 42, 43, 44, and 81 and probable high risk types 53 and 66. The assay was performed according to the manufacturer's protocol. After storage at -20°C, DNA was extracted by cell lysis and centrifugation. DNA amplification was performed in a Perkin-Elmer GeneAmp PCR system 9700 Therma cycler using 5μl of DNA template, 19.25μl of master mixture, and 0.75μl of DNA Taq polymerase. The protocol for amplification was 9 mins of denaturation at 95°C, 40 cycles of denaturation at 95°C for 20s each, 30s of annealing at 55°C, 30s of elongation at 72°C and final extension for 5 minutes at 72°C. This was done alongside positive and negative controls. After amplification the sample was denatured by heating at 95°C for 5 minutes, mixed with 0.5mls pre-warmed hybridisation solution and incubated for 20 minutes, after which a blocking solution was added. This flow-through hybridisation procedure was performed in a sample well atop a Hybrimem HPV-21 membrane containing immobilised probes against which target molecules were directed. Streptavidin-horseradish peroxidase conjugate was added to bind to the biotinylated PCR products. The direct visualisation of the breakdown product (purple precipitate) of the substrate nitroblue tetrazolium-5-bromo-4-chloro3-indolylphosphate was interpreted as positive for the corresponding HPV DNA type as indicated on the schematic diagram of the membrane provided with the test kit.Data analysis: The data collected were analyzed using the Statistical Package for the Social Sciences for Windows version 22.0 . The agreement between the two collection methods were calculated using concordance and discordance rates and the Cohen's kappa statistic. The crude percent agreement between the two techniques was the percentage of samples concordant or discordant for the presence of high risk HPV DNA. The kappa statistic was calculated to determine the level of chance-agreement between the two methods with a kappa value of 0 indicating no agreement better than chance, a value of 1 indicating perfect agreement better than chance, and intermediate values of 0.00-0.20, 0.21-0.40, 0.41-0.60, 0.61-0.80 and > 0.81 indicating poor, fair, moderate, good and excellent agreement respectively. P values of <0.05 were taken as statistically significant.A total of 194 women were recruited for the study. The age range was 23-75 years . The peak age group of respondents was 30 to 50 years, while women above 60 years accounted for the fewest (5%). The mean age of first menstrual period was 18 years, of first sexual exposure was 23.3 years, and of first pregnancy was 25.7 years. The mean parity was 4. Majority of the participants (86%) were in monogamous marriages. Most of the women in this study 76.8% had only one life time sexual partner. Twelve of the 194 (6.2%) women showed positive HPV DNA on provider sampling, while 19 of the respondents (9.8%) showed positive HPV DNA on self-sampling. Multiple HPV genotypes were seen in 1 case (0.5%) and 5 cases (2.6%) of provider collected and self collected samples respectively. The commonest HPV DNA type seen on both provider and self collected samples was HPV 58 . All HPV types seen on provider collected samples were high-risk types, giving a high risk HPV detection rate of 6.2%. Fourteen high-risk HPV types were seen on self sampling, giving a high-risk HPV detection rate of 7.2%. When all the high risk HPV subtypes were considered as a group (group A) against non-high risk HPV/ no HPV as a second group (group B), there were 183 concordant and 11 discordant cases between both sampling methods, i.e. there was an agreement on 7 cases as having high risk HPV, 176 cases as having non-high risk/no HPV and a disagreement on 11 cases as having high risk HPV between both sampling methods. Of the 7 concordant cases, both sampling methods detected the exact same HPV subtype in 6 cases. This translated into a concordance rate of 93.8%, a discordant rate of 6.2%, and an overall moderate agreement between the two methods .Cancer of the cervix continues to be a public health problem in developing countries with higher age standardized incidence rates, life time risk and mortality rates than western societies. The age stand-ardized incidence rate of cervical cancer in West Africa in 2012 was reported to be 29.3/ 100,000 women. This value is high when compared to figures from western countries: 6.6/100, 000 women in the North America and 7.3/100,000 women in the Western Europe. The same is the case for life-time risk of developing cervical cancer (3.2/100 in West Africa compared to 0.7/100 in Western Europe) and the mortality rate  . These fThe detection rate of HPV also depends on the assay method used. The different genotyping tests used in various studies make comparison of results difficult. In one report, self collected vaginal samples had a sensitivity of 70.9% for ≥ CIN 3 compared to 95% for provider collected endocervical samples when assayed using cervista. The sensitivity increased to 94.3% for self collected virginal samples, which was identical to the sensitivity (94.3%) obtained on provider collected sample when the more sensitive PCR based matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was used . PCR basSelf-sampling for HPV DNA, as an alternative to sampling by health care givers, has been well received in some studies. In one report of Hispanic respondents, the overall experience was reported as excellent by 33.7% and very good by 30.8%. Only 2.6% reported a poor of fair experience. The clarity of instructions, ease of use of kit and understanding of results were aspects of the experience the respondents found favourable . A studyThis study found moderate correlation between self and provider collected samples for HPV DNA detection. However due to the small sample size and single centre of study, we recommend larger multicenter studies which would provide more representative results of HPV infection rates in the Nigerian population.Incidence rates of HPV in Ibadan, Irun and Lagos respectively are 26.3%, 14.7% and 11.34%;The commonest HPV types are 35 and 16 in Ibadan, 35 and 52 in Abuja, HPV 52 in Kenya, and HPV 16 and 66 in Chile,the overall experience of self sampling is favourable: it was reported as excellent by 33.7% and very good by 30.8% among Hispanics;The attendance rates for cervical screening increased from 23.2% to 33.2% in Norway when self-sampling was introduced, there is good correlation between self and provider sampling: 85.7% concordance in Canada, 93.8% in India, 93.2% in Seoul and 92% in Uganda.The commonest HPV DNA in this study is HPV 58. This is of epidemiologic significance as it is a high risk type;There was a concordance of 93.8%, and a discordance of 6.2% between both sampling techniques. The concordance is high when compared with other studies;There was an overall moderate agreement between the two methods .