HPV infection causes cervical cancer, a major contributor to morbidity and mortality among low-income Mexican women. Human papillomavirus (HPV) DNA testing is now a primary screening strategy in Mexico’s early cervical cancer detection program (ECDP). Research on Mexican women’s perceptions of HPV and testing is necessary for establishing culturally appropriate protocols and educational materials. Here, we explore perceptions about HPV and HPV-related risk factors among low-income Mexican ECDP participants.We conducted semi-structured interviews with 24 ECDP participants from two primary care health clinics in Michoacán state, Mexico. Interviews addressed women’s understandings of and experiences with HPV and HPV testing. Analysis was inductive and guided by the Health Belief Model with a focus on gender.Women’s confusion about HPV and HPV screening caused emotional distress. They understood HPV to be a serious disease that would always cause severe symptoms, often characterizing it as analogous to HIV or inevitably carcinogenic. Women also attributed it to men’s sexual behaviors, specifically infidelity and poor hygiene. Women described both sexes’ desire for sex as natural but understood men’s negative practices of masculinity, like infidelity, as the causes of women’s HPV infection. Some women believed dirty public bathrooms or heredity could also cause HPV transmission.These results are consistent with prior findings that geographically and economically diverse populations lack clear understandings of the nature, causes, or symptoms of HPV, even among those receiving HPV testing. Our findings also reveal that local cultural discourse relating to masculinity, along with failure to provide sufficient education to low-income and indigenous-language speaking patients, exacerbate women’s negative emotions surrounding HPV testing. While negative emotions did not deter women from seeking testing, they could be ameliorated with better health education and communication. Cervical cancer is one of the main causes of morbidity and mortality among Mexican women, especially low-income women, despite the presence of a national early cervical cancer detection program (ECDP) since the 1970s.  In 2008,Despite the benefits that HPV testing offers as a primary ECDP screening tool, the concept of cervical cancer and screening as virus-related have been found to cause some negative psychosocial effects. Women being tested for or diagnosed with HPV may experience anguish, anxiety, confusion, fear or stigma.–19 TheseSuch perceptions and experiences regarding HPV and other sexually-transmitted infections (STIs) are profoundly shaped by local culture, especially norms and beliefs regarding sexuality and gender.,22 In MeFor these reasons, it is important to assess women’s beliefs about and experiences of HPV in diverse local contexts, in order to understand how they might influence women’s screening experiences. This information is also important for developing educational materials and culturally-sensitive screening protocols, which would increase ECDP use and mitigate any psychosocial harm that screening might pose. Here, we focus on the ECDP experiences of low-income Mexican women in marginalized areas of Michoacán state.All participants provided verbal informed consent, which we employed since low levels of education and literacy in the study population made written consent inappropriate. Consent was obtained through the following procedure. The principal investigator read the letter of informed consent to each potential participant, which included a description of the study including its objectives, potential risks and benefits, the voluntary nature of participation and participants’ ability to withdraw at any time. Potential participants were also given an information card that listed the study title, names, titles and contact information for the ethics board members and study principal investigator, and a statement that the participant could call and ask for information or make comments or complaints. The principal investigator also read this card aloud to potential participants. Women who wished to participate then gave verbal consent, which was audio recorded. As mandated by the local ethics board, this procedure occurred in the presence of at least one witness. This study, including the verbal consent procedure and script followed, was approved by the Mexican National Institute of Public Health Ethics Board (IRB).We conducted semi-structured interviews with women who underwent HPV testing in the ECDP in order to explore their beliefs and perceptions regarding HPV. Participants were ECDP service users drawn from the patient pools of two primary health care centers in the Michoacán State Health Service. These two health centers were selected because they were located in low-income municipalities but served populations that varied ethnically. One center was in the state capital of Morelia, and had a Spanish-speaking, mestizo population. The other center was in the town of Chilchota in Northern Michoacán, which had a large indigenous population composed of Purepecha and Tarasco speakers who had experienced a long history of marginalization from state and economic offerings. We drew From each municipal health care center, we received a list of women who had undergone HPV testing and were given results within the past month. We then verbally invited those women to participate, stopping after 12 women from each site had agreed to participate. Invitations were given in one of three ways: in person at a woman’s next clinical appointment if she had one scheduled during the recruitment period, over the telephone, or at her home if she could not be reached by phone and did not have an appointment scheduled. We explained the purpose of the study, the fact that her participation was voluntary, and that her information would be confidential. We also described the study aims and methods. We then explained the study and collected verbal informed consent from women who wished to participate, as discussed above. Interviews with consenting participants subsequently took place in private rooms at the clinical sites, with the exception of three which were held in private spaces in the homes of Morelia participants for their convenience.The design of the broader study of women’s and healthcare providers’ HPV-related experiences, of which this research forms a part, is described in detail elsewhere. ParticipQuestions collected sociodemographic data and assessed participants’ perceptions and phenomenological experiences of HPV and HPV testing. Question design was guided by the Health Beliefs Model (HBM), which indicates that experiences of vulnerability  and perceptions of severity or seriousness of an event  together influence adoption of preventative behavior . IntervieOur analytic aim was to assess participants’ perceptions and experiences of HPV and HPV testing in the context of local culture, in relationship to the components of the HBM model described above. Data analysis was inductive, meaning that it was guided by the specifics of the data collected rather than a hypothesis based on our theoretical model. The sample size of 24 participants, with 12 from each site, was sufficient for achieving data saturation in a thematic analysis framework, specifically for identifying themes common to multiple participants and discerning whether thematic differences existed between the two sites. We identHere, we provide an overview of participant demographics, then present the key themes that arose in women’s interviews, exemplified with representative quotations. Two main kinds of results emerged from the data, regarding women’s beliefs about the nature and severity of HPV, and their perceptions of HPV’s causes and risks  and HIV (VIH)). These ideas led women who had not seen the Pap test as especially daunting to be more frightened by HPV testing, which they associated with potentially severe outcomes. Women’s perceptions that HPV was very severe related to these beliefs about symptoms, and to the similarity that women perceived between HPV and HIV.Such misunderstandings of HPV led to great stress for many women undergoing screening. Women’s knowledge that the disease was sexually transmitted and their fears that it would inevitably cause distressing symptoms also led participants to believe that many women would not get tested due to embarrassment. For women who were primary speakers of indigenous languages, language barriers added to this confusion and augmented their distress. They especially expressed confusion regarding the basic information they were given, for instance understanding “negative” results to mean harmful results, or not understanding the words “positive” or “negative.” However, as with the woman quoted above who would rather be “hurt” by testing to maintain her long-term health, study participants often said that the emotional discomfort of testing was worth the possibility of arresting terminal illness.In spite of the perceived benefits of testing, confusing messages from health professionals regarding notification procedures and test results made the process emotionally painful for many women. In the ECDP before the introduction of HPV testing, women were notified only if they required follow-up care for abnormal cytology. With HPV testing, women are notified of positive HPV test results which might or might not indicate the presence of pre-cancer. Despite the fact that positive test results are now less closely linked to the presence of pre-cancer, this difference was not communicated and simply receiving results was thus frightening for participants. This led to experiences like that of a woman in Chilchota, who said that she never used to worry about cytology screening but became concerned about her HPV test because of the method of notification of test results. Previously with cytology testing, not receiving paperwork meant a negative result, but when she underwent HPV testing, “the paper arrived.” Since under the prior system receiving “paperwork” meant that one was positive for cervical cancer, participants took positive HPV results to indicate cervical cancer positivity. Women who received positive HPV test results thus experienced distressing confusion regarding the meanings of those test results and the relationship of HPV to cervical cancer. However, the two participants who had recently undergone colposcopy felt that they had received clear explanations about the relationship of HPV positivity to cancer risk. It appears that the explanations provided during colposcopy were more clear and detailed.Women most frequently understood HPV to be transmitted by problematic sexual behavior. They described desire for sex, for both men and women, as a natural need. However, they feared their own needs would put them at risk. They understood this risk to be rooted in the bodies of untrustworthy male partners, viewing men’s negative practices of masculinity, like promiscuity and infidelity, as actions through which men acquired HPV and would pass it on to female partners. Participants also understood poor hygiene to be a cause of HPV risk, in ways that were linked to their understandings of dangerous male sexual behavior. Some believed that there were more sexually transmitted infections around than in previous eras, and attributed this development to men’s general dirtiness, which encompassed both lack of sexual scruples and poor hygiene. Several participants also identified dirty public restrooms as an additional possible HPV vector. They noted this in concert with the risk posed by men’s poor sexual hygiene and infidelity; this implied that rumors that HPV can be transmitted by dirty bathrooms are voiced but seen more as a cover for transmission through male infidelity than a serious, independent risk factor.A few women believed that HPV could be hereditary in addition to sexually transmitted. They often felt concern and sought screening when female family members received positive test results. Since women were not informed that HPV can have a very long incubation period, heritability also seemed like a logical explanation for women who had not had sex for several years, for example while their husbands were working abroad.The overall finding that participants understood that HPV was sexually transmitted but were confused about its symptoms and its relationship to cervical cancer and other STIs mirrors results from diverse world regions and social classes, indicating that low knowledge regarding HPV is a global problem.,35 PriorWe found that study participants shared very similar beliefs about HPV and perceptions of HPV risk regardless of identifying as mestiza or indigenous, other demographic differences such as marital status, educational level and number of children, or whether or not they had received a positive HPV diagnosis. The majority of women participating in the Oportunidades Program and especially the subset of those women participating in Seguro Popular had more contact with health professionals, since they had access to more extensive health care services than others. However,While testing for and diagnosis of sexually transmitted infections can cause emotional pain and stigma worldwide,,41,50,51The women interviewed here underwent HPV testing despite their confusion and negative emotions regarding HPV and the testing experience, and their beliefs that others might be too embarrassed to undergo testing. While this study only included women who had undergone testing and thus did not capture the experiences of women who might have refused it, study participants’ fears about HPV severity facilitated rather than hindered their participation. However, their testing and diagnosis experiences were characterized by negative emotions related both to the gendered ideas discussed above, and poor communication by health professionals. This finding suggests that promotion of HPV testing and cervical cancer screening is not enough; culturally- and linguistically-appropriate education about those issues, as well as clearer communication between health workers and screening participants about these topics, is needed to increase screening among low-income Mexican women by making it a more positive experience.This study’s small sample size and focus on women who had participated in screening and received HPV test results, as well as inclusion only of women who spoke sufficient Spanish, limits the generalizability of its findings. So does the fact that only indigenous participants in one of the two research sites received positive HPV diagnoses. However, these qualitative findings provide important information about the perceptions of HPV and its causes that low-income Mexican women may share. Given that trends in the findings agree with results from diverse populations worldwide, these results support a call for better HPV-related education and doctor-patient communication–especially for people receiving HPV testing–worldwide. They also provide context-specific information that can be useful for improving education and screening protocols for low-income Mexican women.Education regarding the causes of HPV, and the fact that high-risk strains might but do not always inevitably cause cervical cancer in a given individual, is needed worldwide. In Mexico, EDCP protocols should include clear and comprehensive education about HPV in patients’ native languages, and clear communication and open dialogue regarding HPV testing procedures and the meaning of positive test results in relationship to cervical cancer risk and future medical appointments. Mexican sexual health care more broadly should also provide space for patients to discuss fears regarding the relationship of sexually transmitted infections to infidelity, especially in relationship to ongoing cultural debates about masculinity.S1 Table(DOCX)Click here for additional data file.


Laryngeal papillomatosis (LP) is a rare human papillomavirus (HPV) related disease that often requires multiple surgical interventions and residual impairment of voice is almost inevitable. We report the case of a patient with adult onset recurrent LP, showing moderate dysplasia and coinfection with HPV types 6 and 18. The tetravalent HPV vaccine Gardasil was prescribed off label, with the aim of triggering an immunogenic response and consequently reducing the probability of further recurrences. The patient was followed for 9 months with no sign of relapse of his LP. The postexposure use of the anti-HPV vaccine could represent a promising therapeutic agent in established LP. Unfortunately, the potential efficacy of this new therapeutic option in this situation has been suggested only by isolated case reports. Further controlled studies, with a longer follow-up and a larger sample size, are needed to assess efficacy of Gardasil in LP. Laryngeal papillomatosis (LP) is a rare disease with estimated incidence in USA of 4,3/100 000 in children and 1.8/100 000 in adults [In contrast to the low incidence of LP, many studies have shown that HPV DNA can be detected in the upper airway of healthy adults and children , suggestIn children, the infection is generally considered as vertically sexually transmitted from mother to child in the birth canal . In adulDespite its benign nature, LP may significantly affect quality of life as it has a tendency to grow and extend throughout the entire respiratory tract, leading to dysphonia  and dyspnoea . In addition, the role of HPV in malignant transformation is well-described and discussed .For all these reasons, LP is a frustrating and a challenging disease to manage, which may require multiple surgical interventions to remove papillomas and residual impairment of voice is almost inevitable.The ideal therapies aim to maintain airway patency, improve voice quality, and avoid complication.2, argon, and Nd-YAG), and microdebrider, is the preferred mode of treatment but does not prevent lesions from recurring.Surgery, with classic cold knife microsurgery, laser , product from Merck, and Cervarix, product from GlaxoSmithKline, are licensed in more than 100 countries and immunization programs in adolescent girls have been widely diffuse with the intent to prevent cervical cancer .Gardasil is a quadrivalent vaccine made up of recombinant HPV proteins (L1 capsid antigens) from the most common high-risk HPV types (16/18), responsible for 70% of cervical cancer cases and HPV related head and neck cancer, as well as two low-risk HPV types (6/11) which are the causative agents for laryngeal papillomatosis and genital warts. Although the tetravalent vaccine is used to prevent infection by the four subtypes, it has also been used as adjuvant therapy to downregulate the disease in patients with LP.Cervarix is a bivalent vaccine that was FDA approved three years after Gardasil, effective against HPV-16/18. In addition it seems to offer a cross-protection against types 31, 33, and 45, extending its action against 85% oncogenic types.We present a case report of adult onset of LP treated with surgery and immunotherapy based on vaccination with Gardasil.We report on the case of a 48-year-old man, with no relevant comorbidities and no history of smoking or alcohol abuse, sexual transmitted infections, or immune deficiencies.2 laser. After the final surgery an adjuvant treatment with multiple local injections of cidofovir was carried out. This was classified as nonaggressive according to the Lindeberg classification (less than ten operations in 1 year) [In his early forties he developed progressive hoarseness and at the age of 45 he was finally diagnosed with LP. According to the anamnesis data since the diagnosis three surgical interventions were performed: the first one with cold steel and the last ones with CO 1 year) . HistopaFollowing a new severe recurrence, characterized by confluent papillomas, he was admitted to the ENT Department, AOU SS, University of Sassari, Italy. On clinical examination the papillomatosis involved the epiglottis and the false and the true vocal cords bilaterally, resulting in mild dyspnea and severe dysphonia.2 laser. The histology reported papillomatosis with moderate and diffuse dysplasia. Tumor cells exhibited strong and diffuse nuclear and cytoplasmic stains for p16 (NK4a).Surgical debulking was performed using COhttp://www.who.int/biologicals/areas/vaccines/hpv_labnet/en/webcite).The virological analysis was carried out in triplicate at the WHO quality-assured laboratory of the Hygiene and Preventive Medicine Unit, Department of Biomedical Sciences, University of Sassari (A sample of formalin-fixed and paraffin-embedded tissue lesion was selected. DNA was isolated after extraction and purification using Easy-DNA kit . HPV detCoinfection by HPV-6 and HPV-18 was detected. Confirmation was obtained by testing the sample with an “in-house” real-time quantitative TaqMan PCR assay .On the basis of the virological results, three biological samples, collected during the previous surgical interventions, were retrieved and analyzed using the above-mentioned molecular technique. All three were positive for HPV-6.2 laser. In the past cidofovir has been considered an option but the risk of malignant transformation induced by this drug made it not suitable as adjuvant therapy. In addition to this last observation, considering also the failure of the previous attempts to improve the course of disease with antiviral medication, the detection of a viral coinfection with an oncogenic HPV subtype, and the increase in grade of dysplasia over the years, we chose to vaccinate the patient to try to stimulate an immunological response. Moreover the patient showed a large larynx involvement, the most extensive treated in our hospital in the last two decades, inducing us to believe in a necessity to find a good adjuvant therapy to gain a better outcome.Until now in our department the first line treatment has always been surgical, with COThe patient was also tested for anti-HPV antibodies, without positive result, reinforcing the idea that laryngeal HPV infection is not always capable of triggering an adequate immune response . HoweverThe consequent lack of correlation between antibodies level and laryngeal status suggests an inability to produce immune response. Therefore that evidence conferred a little value to the serology test as a screening investigation to detect high risk groups, because even when there is a correspondence with HPV DNA detection, it does not provide any proof of anti-HPV T cell activity, which plays a fundamental role in the immune response against the virus.Considering all of this data the tetravalent HPV vaccine Gardasil was prescribed off label, with the aim of triggering a highly immunogenic response to HPV-6 and HPV-18 and consequently reducing the probability of further recurrences and malignant transformation. A three-dose schedule of vaccination in 1, 3, and 6 months was planned, with the first dose administered 1 month after the surgical intervention.Since the surgical treatment and over a follow-up period of 12 months, no signs of relapse or other mucosal lesions were noted. This represents the longest documented period free of recurrence of our patient.HPV vaccination, currently recommended for noninfected individuals, could represent a promising therapeutic agent in established LP. Unfortunately, the potential efficacy of this adjuvant therapeutic approach has not been assessed in multicenter clinical trials but has been implicated only by few case reports –19.Only Boltezar et al.  and YounWe report a case of adult onset recurrent disease LP. Although nonaggressive, the management of the case was complicated by the coinfection of low risk HPV-6 with oncogenic HPV-18 and thus the risk of malignant transformation, as described in the literature , 23.During 12 months of regular ENT examination, no sign of recurrence or sinister mucosal changes were observed, but a lifelong surveillance should be carried out to detect any early relapses or malignant transformation.More extended studies with longer follow-up and a larger sample size are needed to assess the effectiveness of vaccination in patient with recurrent LP.Another important consideration about the possibility to offer the vaccination in the patients with LP should be suggested by the evidence that this subgroup has less ability compared with the general population to clear this viral infection. Their susceptibility to contract HPV infection in lifetime makes them more exposed to the risk of HPV related cancer, as oropharyngeal cancer, whose rate showed increase in the last decades.From this point of view LP subjects can be considered high risk group, reinforcing the utility of vaccination among them and suggesting a potential benefit from administration of both vaccines.Hopefully the broad spread of HPV vaccines will offer in the future a further decrease of incidence of this disease.


Liquid-state specimen carriers are inadequate for sample transportation in large-scale screening projects in low-resource settings, which necessitates the exploration of novel non-hazardous solid-state alternatives. Studies investigating the feasibility and accuracy of a solid-state human papillomavirus (HPV) sampling medium in combination with different down-stream HPV DNA assays for cervical cancer screening are needed.careHPV™, and Cobas®4800 tests. All the women underwent a rigorous colposcopic evaluation that included using a microbiopsy protocol.We collected two cervical specimens from 396 women, aged 25–65 years, who were enrolled in a cervical cancer screening trial. One sample was stored using DCM preservative solution and the other was applied to a Whatman Indicating FTA Elute® card (FTA card). All specimens were processed using three HPV testing methods, including Hybrid capture 2 (HC2), careHPV™ (83.3 %), and Cobas®4800 (91.7 %) tests. Moreover, the FTA card showed a higher specificity compared to a liquid-based carrier for HC2 , comparable specificity for careHPV™ , but lower specificity for the Cobas®4800 test . Generally, the FTA card-based sampling medium’s accuracy was comparable with that of liquid-based medium for the three HPV testing assays.Compared to the liquid-based carrier, the FTA card demonstrated comparable sensitivity for detecting high grade Cervical Intraepithelial Neoplasia (CIN) using HC2 (91.7 %), FTA cards are a promising sample carrier for cervical cancer screening. With further optimization, it can be utilized for HPV testing in areas of varying economic development. Cervical Cancer is the fourth most common cancer of women worldwide. An estimated 528,000 new cases and 266,000 deaths from cervical cancer occur annually, among which 85 % occur in developing countries. China, a developing country with high cervical cancer incidence 7.5/100,000) and mortality , bears a substantial burden of cervical disease [00,000 anCurrent evidence shows that persistent infection with carcinogenic human papillomavirus (HPV) types is the cause of virtually all cervical cancer . ConsequSolid-state specimen carriers, such as the Whatman Indicating FTA Elute® card , provide an alternative sampling medium for DNA sample collection. FTA cards provide a cost effective room temperature method for collecting, shipping, archiving and processing nucleic acids from a wide variety of biological samples. The cards contain an inert dye that changes from purple to white indicating the location of a clear, colorless sample. Moreover, indicating FTA Elute cards facilitate rapid purification of nucleic acids in less than 30 min per sample and provide DNA in solution for multiple amplification reactions with high stability. There is evidence that genomic DNA stored on FTA cards at room temperature for more than 17.5 years still can be successfully amplified by PCR. FTA cards have been used in forensic science for DNA sample collection and storage for decades, but it has only recently been explored as an alternative to current liquid-based sampling mediums for cervical cancer screening .A few studies have investigated the FTA cards with PCR-based detection methods or a single HPV DNA testing method with high clinical sensitivity and specificity , 9–12. HTherefore, our project sought to investigate the feasibility of using FTA cards as an HPV sampling medium for cervical cancer screening. We also explored the compatibility of FTA cards with the predominant clinical HPV DNA assays.careHPV™ and Hybrid Capture 2 ; the other clinician-collected specimen was tested for HPV16/18/45 E6 protein with the OncoE6™ Cervical test . After sample collection, women were screened by visual inspection with acetic acid (VIA). One to two weeks later, women who tested positive for any of the six screening tests performed  and approximately a 10 % random sample of women who tested negative for all screening tests (screen-negative women) were called back to undergo a second VIA and a rigorous colposcopic evaluation. All colposcopically detected abnormalities (acetowhite lesions) were biopsied. If the colposcopic examination showed no lesion in a quadrant but any of the screen results was positive, a random biopsy was obtained at the squamocolumnar junction in that quadrant at 2, 4, 8, or 10 o’clock. An endocervical curettage (ECC) was performed after the cervical biopsies. Women who were screen negative and were without any colposcopic indications of abnormality did not undergo colposcopically-directed biopsies.Samples were collected among women enrolled in “LCMCCSS”, a large population-based screening trial in rural China, which was a collaboration between the Cancer Institute, Chinese Academy of Medical Sciences (CICAMS) and the Program for Appropriate Technology in Health (PATH) , 14. Elin = 396) was reached. The population included 207 women who were HPV positive, 17 women who were VIA positive (including 10 HPV positive women) and 182 randomly selected normal women. Ethics approval was obtained from the Ethics Committee of Cancer Institute and Hospital, Chinese Academy of Medical Sciences . Written Informed consent was obtained from each subject. One liquid sample was collected by a physician using a Conical Cervical Sampler  and was stored in DCM preservative solution . A second sample was collected by a physician using a new Conical Cervical Sampler and applied to an FTA card, which will be referred to as the “solid sample” in this study. All the samples were stored at room temperature and analyzed in the CICAMS central laboratory. All the women underwent a rigorous colposcopic evaluation that included using a microbiopsy protocol as previously described [We conducted our study among women who returned for colposcopy in LCMCCSS between May and June 2012. Recruitment was stopped when the target sample size . As HC2 and Cobas®4800 test used the same buffer, those tests shared nine punched disks while the careHPV™ assay. Thereafter, PCR amplification was performed to acquire final results for Cobas®4800 test.The six disks and nine disks were transferred into separate 1.5-ml microfuge tubes. Five hundred microliter of sterile water was added to each tube, and the tubes were immediately pulse vortexed for 15 s. Water was removed with a sterile fine-tip pipette. This process was performed twice. Sixty microliter of DCM preservative solution and 90 μl of DEPC water were added to each tube. Both tubes were centrifuged for 30 s and transferred to a heating block at 95 °C for 30 min. During the incubation period, both tubes were centrifuged for 30 s every 10 min to minimize condensation. At the end of heating process, both tubes were pulse vortexed approximately 30 s and centrifuged for 30 s. The eluted DNA was placed into new microcentrifuge tubes and stored at −80 °C until analysis. Finally, 50 μl of the eluate originating from the nine disks was used for HC2 and 25 μl was used for Cobas®4800. Similarly, 50 μl of the eluate from the six disks was used for the www.OpenEpi.com), an open source epidemiologic statistics for public health. Statistical significance was assessed by two-tailed tests with an alpha level of 0.05.The primary objective of this study was to evaluate the sensitivity and specificity of FTA card-based sampling for the detection of CIN2+. According to previous studies, we assumed that the sensitivity of detecting CIN2+ would range from 0.85 to 0.96, the specificity of detecting CIN2+ would range from 0.60 to 0.80, and the CIN2+ prevalence would be approximately 4 %. Given these parameters, we calculated a necessary sample size of 375 with an alpha of 0.05 and 80 % power . AssuminP = 0.005) and a lower smoking rate  than non-participants. Based on their test results, 1392 women were referred for colposcopy, among which 106 (7.6 %) refused to return. The women who returned for colposcopy (n = 1286) had a lower smoking rate  and fewer live births  compared to those who refused to return. For this study, 396 women, with a mean age of 43.7 ± 8.9 years, were recruited from among the women who were referred for colposcopy. All of the subjects were married and the average ages of initiation of sexual activity and menarche were 20.2 ± 2.2 years and 15.5 ± 1.7 years, respectively. More than half of the women (56.5 %) reported primary school as their highest educational level, 33 % reported middle school, and only 10.5 % reported high school or college education. The majority were farmers or housewives (73.5 %), non-smokers (99.3 %), and non-drinkers (86.4 %). Ninety-seven percent (96.9 %) of the women reported having three or less sexual partners, 62.7 % had three or less pregnancies, and 85.4 % had three or fewer live births  refused to participate. The participants had a higher marriage rate , nine as CIN Grade 2 (CIN2), two as CIN Grade 3 (CIN3), and one case as High Grade Cervical Glandular Intraepithelial Neoplasia (HCGIN).Of the 396 samples, 17 were used for careHPV™ (28.8 %) and Cobas®4800 (32.1 %) tests. Compared to the liquid samples, FTA-based samples showed lower HPV positivity for HC2 (22.8 %) and careHPV™ (23.9 %), but a higher HPV positivity for Cobas®4800 test (39.4 %). The FTA card evidenced higher specificity for the HC2 test compared to the liquid medium , but a relatively lower specificity for the Cobas®4800 test . For the careHPV™ test, two sampling methods demonstrated comparable specificity . For the other accuracy parameters, FTA and the conventional DCM sampling medium provided comparable results. Specifically, the FTA card demonstrated a sensitivity identical with that of the DCM medium for HC2 (91.7 %), careHPV™ (83.3 %), and Cobas®4800 test (91.7 %). Moreover, with similar negative predictive value (NPV), the FTA card showed comparable positive predictive values (PPV) for HC2 , careHPV™ , and Cobas®4800  tests.VIA showed a high specificity (95.8 %) for detecting CIN2 or worse (CIN2+) lesions, but a low sensitivity (8.3 %). Only one of 12 CIN2+ cases was identified by VIA. HPV prevalence and the accuracy of detecting CIN2+ lesions for six combinations of two sampling mediums with three HPV testing methods are shown in Table careHPVTM tests failed to identify one and two CIN2+ cases, respectively. The Cobas®4800 test identified all the CIN2+ lesions except for one case of HCGIN. We also calculated the overall percent agreement of three HPV testing methods between two sampling mediums. The results showed a good overall percent agreement of 90.2 % (95%CI: 87.2–93.3 %), 83.7 % (95%CI: 79.9–87.5 %), and 85.6 % (95%CI: 82.0–89.2 %) between FTA card and liquid based sampling methods for HC2, careHPV™, and Cobas®4800 tests, respectively  and in previous studies (careHPV™ vs. HC2: 90.0 % vs. 96.3 %) [The combination of FTA cards with HC2 tests was previously explored in Holland . Our stu 96.3 %) , 20. HowGood agreement between liquid-based media and FTA cards was previously reported for several PCR-based methods, including GP5+/6+, SPF(10) PCR/DEIA /LiPA(25), and real-time PCR assays , 8, 12. Our study is the first to explore the accuracy of FTA cards in combination with the Cobas®4800 test for CIN2+ detection. As the manufacturer did not provide instructions for DCM-based sample detection, CICAMS developed the workflow based on several small pilot studies. Specifically, samples used for PCR amplification from FTA cards had a much higher HPV concentration compared to samples from DCM preservative solution (3.6 % vs. 0.5 %), resulting in a high sensitivity (91.7 %). As a result, a higher HPV positivity and a lower specificity were found for FTA card-based samples compared to DCM samples. Further study is needed to optimize the workflow for Cobas®4800 testing using FTA card-based samples in order to reduce false positives and improve the specificity. Moreover, although this result was comparable with previous studies (70.97 %) of Cobas®4800 test in China  and othecareHPV™ and Cobas®4800 tests and optimized the current workflow for the HC2 test, which provides data for future laboratory investigations. Moreover, this study differs from previous reports which used PreservCyt  medium for a single test; this study is the first to compare the accuracy of the FTA card and DCM preservative solution combined with three clinical HPV DNA tests in cervical cancer screening. This study has some limitations that must be addressed as well. First, our study was based on a referral population that was triaged for colposcopy. A higher HPV prevalence was found in this group compared to the general population, which had an impact on the accuracy parameters for detecting CIN2+ lesions. Second, this study is a hospital-based pilot study with a relatively small sample size. We therefore lacked statistical power to assess test characteristics among sub-groups. Third, the Cobas®4800 test workflow requires further optimization for future large-scale studies. Fourth, we did not consider the sequence of sample collection. Taking two samples with a short time interval might have a negative impact on the accuracy of the second sample. However, the second sample, which was collected on FTA cards, showed comparable accuracy for detecting CIN2+. This indicates that FTA cards have a robust capacity for capturing samples. Future studies should further evaluate the accuracy of these two sampling media in a random sampling sequence to avoid any potential bias. Fifth, we made a parallel comparison for three HPV testing methods on a single FTA card, which may result in insufficient distribution of sample for each test. Additionally, reproducibility studies were not feasible in this study. Finally, there are several types of cervical cytological sampling brushes, but this study only used a conical cervical sampler for sample collection. Future comparisons of different cytological samplers need to be investigated to identify the samplers that provide the highest degree of accuracy.This is the first report of a study comparing three downstream HPV testing methods with clinical implications by FTA cards and conventional liquid based sampling media. This study explored experimental workflows for the careHPV™ could be used for cervical cancer screening in remote areas, and FTA cards in combination with HC2 or Cobas®4800 could be used in metropolitan areas. Our previous study demonstrated that the agreement between self-collected and clinician-collected specimens on FTA cards was very good and the acceptability of FTA card-based self-collection was reasonably high [This work also draws a promising picture for future large-scale cervical cancer screening projects. As dried material on a solid carrier is neither hazardous nor contagious, applying genital self-samples on FTA cards can solve storage and transportation problems encountered in developing regions. With further optimization and automation of the experimental procedures, FTA cards in combination with bly high , 6. Therbly high .Our study provided clinical evidence for evaluating the accuracy of FTA card-based sampling as an alternative to the liquid-based sampling in cervical cancer screening. With further optimization of the experimental workflow, FTA cards can be paired with different HPV DNA testing methods suitable a variety of economic conditions, thereby providing new possibilities for large-scale population-based cervical cancer screening.


A retrospective observational study was performed based on individual data from two databases. A total of 162 females aged between 20 and 65 years from Uppsala County, Sweden with CIS and an HPV test conducted between 2010 and 2011, preceding or concomitant to CIS diagnosis, were included. Females with cervical squamous cell carcinoma  were used for comparison. In total, 96% (n=156) of females with CIS were positive for high-risk HPV; HPV16 was the most prevalent (44.5%), followed by HPV33/52/58 (19.5%), HPV31 (13.1%) and HPV18/45 (9.5%). HPV16 was most frequently detected in females with CIS aged between 20 and 29 years (73.6%) and least frequently detected in those aged between 50 and 65 years (33.3%), with a statistically significant age-specific difference (P=0.001). Among the HPV16-positive females, multiple infections were most frequent in the younger age groups. The prevalence of HPV16 in females with CIS decreased with age, whereas a high prevalence of HPV16 remained in females with SCC. These results may indicate that HPV16 has increased oncogenic potential in older females.Human papillomavirus (HPV) 16 is the dominant cofactor in cervical cancer development. The present report investigated the age-specific prevalence of HPV16 in cervical carcinoma  Persistent infection with high-risk human papillomavirus (HPV) types has a critical aetiological role in the development of cervical intraepithelial neoplasia and cervical cancer . The proet al (in situ (CIS) followed by HPV33/52/58 in 24%, HPV31 in 13% and HPV18/45 in 12%  in Sweden occurred in postmenopausal females aged ≥50 years  and 86% The purpose of this study was to identify the prevalence of HPV16 in females with CIS in the population-based organised cervical screening programme in Uppsala County, Sweden, and to describe the age-specific prevalence of multiple HPV infections, with a focus on older females. The prevalence of HPV16 in females with CIS was also compared with that in females with SCC.VIR multiplex real-time polymerase chain reaction (PCR) assay. Females with persistent ASCUS/low-grade SIL and high-risk HPV infection at the three-month follow-up, as well as females with high-grade SIL, are referred for a complete clinical work-up, including a pelvic examination, colposcopy with directed biopsies of suspicious areas and repeat cytology.The population-based organised cervical screening programme in Uppsala County, Sweden screens females aged between 25 and 60 years. Within the framework of the programme, ~70,000 females are screened by conventional cytology each year by midwives in family planning centres. Females with abnormal cytological findings, using atypical squamous cells of undetermined significance (ASCUS) as a cut-off, are referred for further investigation. Females with low-grade cytological abnormalities [such as ASCUS and low-grade squamous intraepithelial lesions (SIL)] undergo supplementary HPV testing and have a new cytological sample taken following a mean follow-up time of approximately three months, at which time two cervical brushes  are used to collect cell samples from the ecto- and endocervix. One brush is spread on a slide for Papanicolaou staining, while the other is applied to a FTA elute card  ,14. HPV The SymPathy database  contains information on results obtained at the Department of Clinical Pathology and Cytology of Uppsala University Hospital . Data therein are identified by specific topography and morphology codes . The database of the organised population-based cervical cancer screening programme of Uppsala County, Sweden includes cytological, HPV and histological results from all laboratories in the county. The cytological and histological samples from the screening programme are stored at the Department of Clinical Pathology and Cytology of Uppsala University Hospital , where all cytological, HPV and histological testing for the screening programme is performed. Detailed descriptions of the database searches for HPV status and HPV screening analysis results have been published .Briefly, a 3-mm Ø Harris micro punch  is used to excise sections from the FTA cards using the BSD 600  semi-automatic punch robot. Four punches from each card are then transferred to a single well in a 96-well plate and washed by vortexing three times for 5 sec in 200 μl distilled water. The water is then carefully removed using a pipette. DNA elution is performed in 50 μl of distilled water at 95°C for 30 min in a heating block (with a heated lid). A total of 3 μl of the DNA extract is used as a template in each real-time PCR . HPV typFemales aged between 20 and 65 years with a histopathological diagnosis of CIS determined between 2010 and 2011 were identified in the Sympathy database. Previous cytological, HPV and histological results for these females were taken from the database of the cervical cancer screening programme of Uppsala County, Sweden. The study population was then restricted to females with a preceding or concomitantly performed HPV test, separated by age group and further restricted to females who tested positive for HPV.Older females with SCC were used for comparison. Females aged between 50 and 95 years with a diagnosis of SCC determined between 2008 and 2011, and a preceding or concomitant HPV test were identified and included in the study.2 (for n<5) were used to compare proportions.Data were analysed using the Statistica 6.1 software . Pearson’s and Yates’ corrected χThe present study was approved by the Uppsala University Ethics Review Board , which determined that informed consent from the participants was not required, and was conducted according to the principles expressed in the Declaration of Helsinki.Of the 205 females with CIS identified in the database, 162 (79%) had a previous or concomitant HPV test recorded in the database of the organised cervical cancer screening programme. Of those, 156 (96%) tested positive for HPV and were included in the final analyses. In addition, 35 females with SCC were also included for comparison. The mean age of females with CIS was 37 years,  and the mean age of females with SCC was 66 years .2). Whereas HPV18/45 was only identified in 19 (9.5%) females of the study population . HPV33/52/58 and HPV31 were also more common than HPV18. HPV33/52/58 was found in 13.1% of females aged between 20 and 29 years and in 30% of females aged ≥50 years, and the HPV33/52/58 types were as common as HPV16 in females aged ≥50 years (30.4%) . Age-specific HPV16 prevalence ranged from 73.6% (28/38) in females aged between 20 and 29 years to 33.3% (7/21) in females aged between 50 and 65 years  . The imp (30.4%) .Multiple infections were common in this study population, but more so among the younger females. Overall, 38.9% of females harboured more than one HPV type, and 26.3% of females aged between 20 and 29 years harboured more than one high-risk HPV type, compared with 9.6% of females aged between 30 and 39 years, and 3.0% of females ages ≥40 years. In HPV16-positive females, multiple infections were also more prevalent in younger females; 35.6% of females aged between 20 and 29 years, compared with ~7.0% in females aged ≥40 years .In total, 30 (85.7%) females with SCC were positive for high-risk HPV types. HPV16 predominated and occurred in 19 (63.3%) females with SCC, followed by HPV18/45 in six (20.0%) females. However, multiple HPV infections were not recorded in any females with SCC .The biopsies of the six females with CIS and a negative HPV test were re-evaluated, but the diagnosis was not altered. The biopsies of the five females with SCC and a negative HPV test were also re-evaluated, but in the majority of cases the biopsies were small, and it was not possible to determine whether the tumour was of cervical origin, or constituted local metastases of a tumour with another primary site.This study provides an overview of the HPV type distribution, and the prevalence of multiple infections encountered among females of all ages with CIS detected in a population- and cytology-based cervical cancer screening programme, as well as among older females with SCC. The prevalence of HPV16 was found to vary with age in females with CIS, with a significant decreasing prevalence in older females. Other studies have identified the same pattern, showing HPV16 as the dominant oncogenic HPV type in CIS ,15. In fA combination of organised and opportunistic screening has reduced the incidence of SCC substantially during the last decades in Sweden ,17. HoweIn conclusion, HPV testing exhibits a considerably higher sensitivity to detect CIS compared with conventional cytology , particu


CCR2-64I (rs1799864) andCCR5-Δ32(rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) ina Brazilian population. The genotyping of 139 women with cervical lesions and 151women without cervical lesions for the CCR2-64I andCCR5-Δ32 polymorphisms were performed using polymerase chainreaction-restriction fragment length polymorphism. The individuals carryingheterozygous or homozygous genotypes (GA+AA) for CCR2-64Ipolymorphisms seem to be at lower risk for cervical lesion . The same was observed for the A allele , while noassociation was detected (p > 0.05) with CCR5-Δ32 polymorphism.Regarding the human papillomavirus (HPV) type, patients carrying theCCR2-64Ipolymorphism were protected against infection by HPV type16 . In summary, our study showed a protective effectofCCR2-64I rs1799864 polymorphism against the development ofcervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.Polymorphisms in chemokine receptors play an important role in the progression ofcervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examinedthe association of  Infections by oncogenic types of human papillomavirus (HPV) are found in 99% of women withcervical cancer (CC) de .TherefoThe loss of cell cycle control mechanism, infiltration of leukocyte, and of otherimmunocompetent cells, as well as the altered expression of immune response genes, havebeen considered critical in neoplasms cervical pathogenesis and progression for CC .Among the gene products present into cervical mucosa, chemokines and their receptor haveshown a key role in immunity against cervical tumours .ChemokiCCR5 gene) leads to decreased expression and dysfunction of CCR5receptor (CCR5-Δ32 (rs333) gene have reduced risk for asthma and early-onsetmyocardial infarction, attenuation of severity in rheumatoid arthritis, and slower acquiredimmune deficiency syndrome (AIDS) progression (Chemokine receptor (CCR)5 is the major receptor for the chemokine and their ligands are themacrophage inflammatory protein (MIP)-1α/chemokine ligand (CCL)3, MIP-1β/CCL4, andregulated on activation, normal T cell expressed and secreted/CCL5 . Polymorreceptor . Studiesgression . In addigression .CCR2-64I, rs1799864) of CCR2 gene that encodesisoleucine (ATC) instead of valine (GTC) has been widely studied, and there are reports ofassociation between this polymorphism and the protective effect in the progression ofinflammatory diseases such as multiple sclerosis -1,being associated with carcinogenesis and angiogenesis . The sinclerosis , carotidclerosis , and in clerosis . Howeverd so far .CCR2-64I and CCR5-Δ32 polymorphisms withdevelopment of cervical intraepithelial neoplasia (CIN) or CC in women infected by HPV fromNortheast Region of Brazil.Therefore, the aim of this study was to analyse the associationforPopulation - The present study was a hospital-based cross-sectionalprospective one carried out in the outpatient clinics of the Lower Genital TractPathology Clinic at the Women’s Healthcare Center of the Prof Fernando FigueiraInstitute of Integrated Medicine, Recife, state of Pernambuco, Brazil. Patients wereselected by spontaneous demand from January 2009 until 2011 and the study populationconsisted of 290 sexually active women ranging between 16-75 years old. Information wascollected from all women pertaining to their age, smoking, alcohol consumption, numberof offspring, number of sexual partners, and age at first coitus. The inclusion criteriawas as follows: women with oncotic cytology submitted to Papanicolaou test according to Bethesda System terminology  buffersolution (Tris-HCl 10 mM and EDTA 1 mM pH 8.0) and conserved at -20ºC untilanalysis.DNA extraction - Genomic DNA extraction was performed from 300 µL ofvaginal fluid from each study subject, following the manufacturer’s instructions of thekit Wizard® Genomic DNA Purification . The analyses sampleswere executed the Laboratory of Genetics, Biochemistry, and DNA Sequencing at RuralFederal University of Pernambuco.HPV detection and typing - Amplification of human β-globin gene segmentwas used as an internal control for DNA quality and samples negative for this assay wereexcluded from analysis. Then, our samples were tested for HPV presence using MY09/11,GP05+ and GP06+ consensus primers by polymerase chain reaction (PCR) (on (PCR) . The typon (PCR) .Analysis of the CCR2-64I polymorphism - TheCCR2-64Ipolymorphism was analysed through PCR followed by restriction fragment lengthpolymorphism .morphism . DNA wasAnalysis of the CCR5-Δ32 polymorphism - TheCCR5-Δ32polymorphism was analysed through PCR .ough PCR  using prSequencing - A total of 20% of the all samples (randomly chosen) wassubmitted to bidirectional sequencing in order to double-check the genotyping results for each polymorphism . The influence of each polymorphism on the riskfor development of (pre) neoplastic cervical disease was estimated by odds ratio (OR)and a 95% confidence interval (CI). Allele frequencies were estimated by directcounting. Comparison between genotypic frequencies of patients and control groups wasperformed by chi-square test and Fisher’s exact test was used to compare the allelefrequencies in contingency tables.For identification of relevant risk factors, a logistic regression analysis was carriedout (comparing HPV-positive women with history of lesions with HPV-positive women withno history of lesions or CC). This modelled the influence of genetic polymorphisms, HPV16 single infection or multiple HPV strains co-infection, smoking and alcoholconsumption on the risk of developing high-grade squamous intraepithelial lesions. TheOR and their respective 95% CI were determined. The R software v.3.0.2 (R-project.org/)was used to perform the regression analysis. Power analysis was performed throughG*Power software v.3.1.9.2 .All p-vWithin the 290 HPV positive enrolled women, 139 had cervical lesions (CIN or CC) (HPV+L)while the 151 had no cervical lesions (HPV+). When considering the prevalence oftype-specific HR-HPV infection, we observed that 38.8% of the patients had HPV 16, 22.3%HPV 18, 2.9% HPV 31, 3.6% HPV 33, and 14.4% other HPV types. Furthermore, the presenceof co-infection by HPV types 16/18 was found in 18% of the patients . MoreoveAmong the 139 HPV+L, five risk factors for cervical lesions were analysed: smoking,alcohol consumption, number of offspring, number of sexual partners, and age at firstcoitus. With the 139 HPV+L, 32.37% (45/139) reported smoking, 60.03% (89/139) alcoholconsumption, 28.78% (40/139) number of offspring > 3, 20.17% (24/119) had number ofsexual partners > 4, and 59.71% (83/139) had first coitus with ≤ 16 years old.CCR2-64I and CCR5-Δ32polymorphismsgenotypes in 139 women with cervical lesions (CIN or CC) and 151 HPV+ were according tothe Hardy-Weinberg equilibrium. A significant difference in the distribution ofCCR2-64I polymorphism between HPV+ L and HPV+ was observed using adominant genetic model , being the variant carrier (GA+AA)associated with protection to cervical lesions  (The distribution of  lesions . When co 0.3943) .CCR2-64Ipolymorphism and susceptibility to HPV 16 infection   ; theCCRCCR2-64I andCCR5-Δ32 polymorphisms with age, smoking, alcohol consumption,number of offspring, number of sexual partners, or age at first coitus (p >0.05).Patients clinical features are shown in CCR2-64IandCCR5-Δ32 polymorphisms and the presence of cervical lesions (CIN orCC) in HPV infected women from Northeast Region of Brazil.In this study, we examined the possible association between CCR5 and CCR2 have been associated withcarcinogenesis and angiogenesis ; CCR2-64I can be linked with decreasedmacrophages recruitment in the process of tumour angiogenesis, which could be a keyduring progression of cervical neoplasia to CC. However, Chatterje et al. (2010)proposed that the raised attraction of the macrophages through the increased expressionof MCP-1 could be auxiliary in the process of destruction or progression of tumour.These discordant findings reinforce the possibility of several factors associated withthe multifactorial neoplastic development, including the difference in ethnic origin ofthe populations studied, sample sizes, and the low percentage of the mutant allele(CCR2-64I).The clerosis , carotidclerosis , breast clerosis , AIDS prclerosis , and CINclerosis . Our resCCR5-Δ32 is involved in slower AIDS progression. However, to clarify the role of this genotype in both HPV infection andprogression for cervical lesion, the CCR5-Δ32 polymorphism should betested in other populations from different ethnic background.Studies reported that gression, in decrgression, and in gression. To our Regarding to prevalence of HPV infection, we found that the more frequent types were HPV16 and 18 . The prevalence of HPV 31 was 2.8% and HPV 33was 3.5%. Previous reports have shown that concomitant or sequential detection of more than oneHPV type it is associated with different stages of cervical lesions . Our resCCR2-64I polymorphism with the susceptibilityof infection by HPV type 16 (OR = 0.35) was observed, suggesting a possible relation ofthis genetic variant with protection to HPV-16 infection.A protector effect of the CCR2-64I and CCR5-Δ32 polymorphisms. However,no significant difference was observed (p > 0.005). The frequenciesofCCR2 and CCR5 variants alleles were in agreementwith a previous study in different diseases and ethnic groups (We also compared clinical features from patients with genotypic distribution ofbothc groups .CCR2-64I polymorphism is associatedwith the protective effect to development of cervical lesions as well as in theprotection to HPV 16 infection.Thus, our data suggest that the


ZNF582) (ZNF582m) was reported to be highly associated with cervical cancer. In this study, we compared the performance of ZNF582m detection and HPV-HR genotyping in the triage of cervical atypical squamous cell of undetermined significance (ASC-US) and atypical squamous cell - cannot exclude a high-grade lesion (ASC-H).The interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR). Recently, methylated zinc finger protein 582  assay that combines degenerate E6/E7 consensus primers and HPV type-specific primers.Two hundred and forty-two subjects with equivocal papanicolaou smear (Pap smear) results were recruited in this hospital-based and case-controlled study. The residual cervical cells in liquid-based cytological test (LBC) containers were used for genomic DNA extraction and then for ZNF582m and the risk of cervical intraepithelial neoplasia grade 3 or higher : 7.73 to 31.18). The sensitivity and specificity of ZNF582m for women with CIN3+ were 82.43% and 76.79%, respectively. High sensitivity (99.33%) but low specificity (38.76%) was observed for HPV-HR. When combining both positive results of ZNF582m and HPV-HR, the sensitivity and specificity were 82.43% and 81.55%, respectively. The sensitivity and specificity of ZNF582m or HPV-16/18 were 89.19% and 70.24%, respectively. However, the sensitivity and specificity of ZNF582m combined with HPV-16/18 (both ZNF582m and HPV-16/18 positive results) were 59.46% and 94.64%, respectively.Significant associations were observed between ZNF582m provides a promising triage tool for women with ASC.ZNF582m and HPV-16/18 genotyping was proposed. This strategy could reduce the number of patients referred for colposcopic examination and thus provide a feasible follow-up solution in the regions where colposcopy is not readily available. This strategy could also prevent women from experiencing unnecessary anxiety caused by HPV-HR.To effectively manage ASC patients, a new strategy co-testing for  Cervical cancer is the fourth most common cancer that affects women worldwide. In China, approximately 58,000 new cases and 20,000 deaths were documented in 2005 -5. ScreeCOL25A1 and KATNAL2, which showed increased methylation during progression in high-grade cervical intraepithelial neoplasia (CIN) lesions, to screen for the early detection of cervical cancer [ZNF582) possesses great clinical potential [ZNF582 is located on chromosome 19q13.43, which contains one KRAB-A-B domain and nine zinc-finger motifs. A recent study of acute myeloid leukemia revealed that aberrant methylation of ZNF582 is consistent in different disease subtypes, especially in the promoter region. Although the biological function of ZNF582 is not yet well characterized, a group in Taiwan found similar associations for cervical cancer and adenocarcinoma with ZNF582 using cervical scrapings. Recently, one study also revealed that hypermethylated ZNF582 (methylated ZNF582 (ZNF582m)) performs well as a marker for detecting grade III or higher cervical intraepithelial neoplasia (CIN3+) with 73% sensitivity and 71% specificity for women with low-grade squamous intraepithelial lesions [The mechanisms of pathogenesis in cervical cancer are not entirely clear. Hypermethylation of CpG sites in promoter regions silences the transcription of tumor suppressor genes and is an early and crucial event in cancer development -13. Inacl cancer ,15. The l cancer -19, and l cancer . Among totential . ZNF582  lesions -23.ZNF582 testing (ZNF582m) for the triage of ASC-US and ASC-H cervical scrapings and compared the results with those of HPV DNA testing.In the present study, we examined the performance of methylated The study was approved by the Department of Clinical Pharmacology, Xiangya Hospital, Central South University, People’s Republic of China. During the period between December 2011 and April 2013, women referred for a colposcopic examination were invited to participate in the study. The eligible women were sexually active, scheduled to undergo a liquid-based cytology test (LBC) within a week, not pregnant, possessed an intact uterus, and no history of treatment for CIN or cervical cancer. The exclusion criteria included women who had a history of cancer related to the reproductive tract, underwent therapy for cervical lesions, received the HPV vaccination, or were currently pregnant. Standard guidelines for the management and treatment of cervical neoplasia were followed in all subjects . The cytAfter signing the informed consent, patients immediately received the colposcopic examination and biopsy. The follow-up clinical information was collected. When the biopsy results revealed moderate to severe dysplasia or worse, patients underwent conization or major surgery. The final diagnosis was based on the results of tissue-proven pathology, rather than on the LBC results. To ensure the quality control of the diagnosis, two expert cytologists and two pathologists independently read or reviewed the cytology and histology slides, respectively. All of the patient recruitment and clinical information collection processes were periodically monitored and good clinical practice (GCP) guidelines were followed.ZNF582. All of the molecular tests were performed at the Institute of Clinical Pharmacology, Hunan Key Laboratory of Pharmacogenetics, China, following GLP guidelines. The cells were centrifuged and stored in phosphate-buffered saline (PBS) solution at −20°C from the day of collection, and the assays were finished within 6 months. Genomic DNA (gDNA) was extracted from the collected cells with a QIAamp DNA Mini Kit  according to the manufacturer’s protocol. A BioSpec-nano spectrophotometer  was used to quantify the amount of extracted DNA.All of the patients had LBC testing before receiving the colposcopic examination. After the informed consent was signed, the LBC containers  were picked up from the Cytology Department, and the residual cervical cells were used for HPV typing and detection of methylated For the DNA methylation tests, quantitative methylation-specific PCR (QMSP) using TaqMan-based technologies was performed using the Lightcycler LC480 real-time PCR system . Briefly, 500 ng of gDNA was subjected to bisulfite conversion using the EZ DNA Methylation-Gold™ Kit , according to the manufacturer’s recommendations.ZNF582 were then determined using the qPCR kits developed by iStat Biomedical located in Taiwan. The type II collagen gene (COL2A) was designed as the internal reference and tested with each specimen. The crossing point (Cp) value for COL2A, which is also the validity indicator of the test, should not be larger than 35. The PCR cycling conditions consisted of an initial incubation at 95°C for 10 min, followed by 50 cycles of denaturation at 95°C for 10 s, and annealing/extension at 60°C for 40 s. Fluorescence data were collected during the annealing/extension step for determination of the Cp. For each sample, two Cp values were obtained: one from the target gene and another from COL2A. The DNA methylation level was subsequently determined from the difference between the two Cp values (delta Cp = CpZNF582 - CpCOL2A). ZNF582 was deemed to be hypermethylated or positive if the delta Cp was smaller than 13 . The registered CaSki and A375 cancer cell lines were used as the methylation and no-methylation controls to ensure the quality of bisulfite conversion and qPCR processes. Representative positive and negative QMSP results are shown in Figure The methylation levels of High-risk human papillomavirus (HPV-HR) typing was performed using a nested multiplex PCR (NMPCR) assay that combines degenerate E6/E7 consensus primers and type-specific primers. The identification of HPV-HR was achieved by determining the size of the nested PCR amplification product .ZNF582m were generated from the first 100 subjects, including 50 subjects with normal Pap test results and 50 subjects with abnormal Pap test results. Receiver operating characteristic (ROC) curves were generated, and the area under the ROC curve (AUC) was calculated for the detection of the CIN3+ lesions.The cut-off values for ZNF582 or HPV typing in different combinations. Fisher’s exact test was used for the small number data, for example, age under 30 group analysis. For the final analysis, the sensitivity and specificity with a 95% confidence interval (CI) for grade CIN3 lesions or worse were obtained. All differences were considered two-sided and statistically significant at P < 0.05.SAS software (version 9.2) was used for all statistical analyses . The chi-square test was used to analyze the status of methylated Establishment of routine Pap screening has resulted in a significant decrease in cervical cancer deaths. In China, ASC, including ASC-US and ASC-H, were reported in approximately 10% of papanicolaou smear (Pap smears) . The larBecause of the high sensitivity, triage of ASC-US with HPV-HR has been suggested in the cervical screening guidelines. This strategy provides the advantage of missing fewer cases of the severe disease state . HoweverN = 74), HPV 16 (55.4%) and HPV 52 (31.1%) were the most common types, followed by HPV 18 (13.5%), HPV 58 (12.2%), and HPV 33 (9.4%). Among the high percentage of HPV 16 and/or 52 in the CIN3 lesions, the sensitivity, specificity, and accuracy are 74.32%, 73.81%, and 73.97%, respectively. Therefore, the combination of Pap smear and HPV-16/18 typing is not sufficiently accurate for cervical cancer triage in this study.In 2012, the American Cancer Society proposed new screening guidelines that included using age-adjusted screening methodologies as well as cytological and HPV co-testing. HPV 16 and 18 typing has been recommended for HPV-HR-positive women . HoweverZNF582m has been found to possess clinical potential for the detection of cancer. For example, the frequency of methylated ZNF582 detection was 100% in SCC tissue. Clinically, ZNF582m has demonstrated 70% sensitivity and 82% specificity for CIN3+ lesions in a Taiwanese case-control cohort. For triage of LSIL, ZNF582m also has good sensitivity and specificity with values of 73% and 71%, respectively. Moreover, the ZNF582m was reported at a high methylation rate in cervical adenocarcinoma, which has been difficult to diagnose.CADM1/MAL, WT1, and other methylation genes have been reported [ZNF582m and HPV-16/18, in which the patients negative for ZNF582m undergo HPV-16/18 genotyping. As mentioned, the sensitivity, specificity, and accuracy of the co-test are 89.19%, 70.24%, and 76.03%, respectively. The colposcopy referral rate of the co-test is 47.9%, which is 25.2% less than that of HPV-HR alone. Although eight cases of CIN3+ (10.81%) were missed, nearly 100% of the women with SCC were successfully diagnosed and treated  observed. Although it is known that heterogeneous etiology for cancers and the equivocal nature of ASC, the significance of in situ (CIS) in 6 (2.47%) patients, and invasive carcinoma in 29 (11.98%) patients. Thus, 30.6% (74 of 242) of the total patients had a histological finding of CIN3 or worse (CIN3+). Specifically, 22.1% and 70% of the patients had CIN3+ in the ASC-US and ASC-H groups, respectively.From November 2011 to March 2013, 242 women were scheduled for colposcopic examinations. The characteristics of the subjects are shown in Table ZNF582m. The sensitivity, specificity, and accuracy of ZNF582m in the identification of CIN3+ lesion were 82.43%, 76.79%, and 78.51%, respectively. However, when the CIN2 or more severe neoplasms were targeted, the sensitivity and accuracy of ZNF582m was decreased to 67.27% and 74.38%, respectively, but the specificity was increased to 80.30%. Among women less than 30 years old with ASC, the sensitivity, specificity, and accuracy of ZNF582m in the identification of a CIN3+ lesion were 75.00%, 81.48%, and 80.65%, respectively. For postmenopausal women with ASC, the sensitivity, specificity, and accuracy were 88.89%, 69.77%, and 75.41%, respectively.The distribution of the molecular tests for different levels of disease severity is listed in Table The HPV typing indicated that 177 (73.14%) patients were positive for HPV-HR. The most common types of HPV were HPV 16 (23.14%), HPV 52 (23.14%), HPV 58 (14.88%), HPV 68 (7.85%), HPV 33 (7.44%), and HPV 18 (6.05%). The sensitivity and specificity of HPV-HR in identifying CIN3+ were 99.33% and 38.76%, respectively. The percentage of patients who were positive for HPV-16/18 and who had CIN3+ was 66.22% (49 of 74). The prevalence of HPV-16/18 in patients whose tissue was pathologically normal, CIN1, CIN2, CIN3/CIS, or squamous cell carcinoma (SCC) was 7.34%, 17.39%, 22.22%, 55.56%, and 82.76%, respectively.ZNF582m combined with different HPV genotyping assays are interesting. The sensitivity of ZNF582m combined with HPV-HR, HPV-16/18/52/58, or HPV-16/18 was 99.33%, 95.98%, or 89.19%, respectively, but the specificity was increased with values of 34.02%, 48.81%, and 70.24%, respectively.The results for ZNF582m alone, HPV-HR alone, and ZNF582m, or HPV-16/18 are 41%, 73.1%, and 47.9% respectively  from colposcopy referral (when compared with HPV-HR), yet none of the SCC cases would be missed  illustrated in a flowchart. The present study did have some limitations. The subjects were recruited at the colposcopic examination room. Other limitations include using a smaller sample sizes and a lack of extensive long-term follow-up information. However, our results suggest that testing for ZNF582m followed by HP-V16/18 testing could help to reduce the number of patients with ASC being referred for colposcopy. This could reduce the logistics of obtaining follow-up care, especially in regions where colposcopy is not easily available, which could have a significant impact on public health.To the best of our knowledge, the present study is the first to discuss the involvement of


Validated measures that can accurate describe young adults’ HPV vaccination attitudes and how these relate to vaccination intention and receipt are needed for developing interventions to improve low HPV vaccination levels. The Carolina HPV Immunization Attitudes Scale (CHIAS) is a validated measure of these outcomes that was originally designed for parents.To assess the performance of the CHIAS among young adult women using an exploratory factor analysis.A convenience sample of 139 young adult women (age 18–26 years) were given the CHIAS measure at baseline. Factor analysis was used to determine attitudinal factor groupings and the association of these factors with HPV vaccination intention. A 6-month follow up assessment examined the stability of the CHIAS over time and the association of baseline vaccine factors with vaccine receipt.Five factors loaded on to the CHIAS in young adults - “Barriers,” “Harms,” “Effectiveness,” “Risk Denial” and “Uncertainty,” - which was similar to the factor loadings of CHIAS for parents. “Harms” was the factor most consistently associated with vaccination intention at all time points assessed. Only 5 women had received or made an appointment to receive the vaccine at the 6-month follow-up.In terms of categorizing HPV vaccination attitudes, the CHIAS appears to have similar performance among young adults as in parents. However, additional studies are needed to assess the utility of the CHIAS for predicting HPV vaccine receipt among the young adult population. Vaccines against human papillomavirus (HPV) represent a remarkable opportunity for the primary prevention of cervical cancer and other HPV-related diseases. Despite these health benefits, HPV vaccination among young adults in the U.S. is significantly lower than national goals. Interventions to improve HPV vaccine utilization among young women have been hindered by limited understanding of the factors that influence vaccine acceptability, intention, and ultimately vaccine utilization among this population. The CHIAS, developed by McRee et al, was originally evaluated among a regional sample of parents in North Carolina. Having a similarly robust, standardized measure of HPV vaccination attitudes for young women would be useful for developing and examining interventions to improve HPV vaccine uptake among this population. Therefore, the goal of this study was to examine the factor structure of the CHIAS when applied to a sample of young adult women. The specific objectives were: 1) to compare the factor structure that results from young women’s use of the CHIAS to that reported previously for parents, and 2) to evaluate the association between the CHIAS factors and young women’s HPV vaccination intention and utilization.We conducted a cross sectional survey of 139 college-aged women that was implemented from October 11, 2011 to November 1, 2012. This survey was part of a larger study aimed at evaluating the longitudinal impact of different educational materials on HPV vaccination intention and receipt , and on hormonal stress responses to those materials (manuscript in preparation). The focus of this manuscript is on responses to CHIAS items specifically.This study was approved by the Institutional Review Board at the University of Michigan. Written informed consent was received from all participants.Participants were recruited via a Psychology participant pool and flyers posted on campus and in the local town advertising a study about HPV vaccines. Eligibility criteria for participation included being a female aged 18–26 years and not yet having received any doses in the HPV vaccine series. Upon arriving to the study lab and providing informed consent, participants received a paper version of the baseline survey to complete in a private cubicle in the lab. Follow-up surveys were emailed to participants and administered via Qualtrics with up to 2 prompts for non-respondents.HPV vaccination is not really necessary because Pap smears can be done to make sure cervical cancer doesn't develop”). Other parents in my community are getting their daughters vaccinated” becomes “My friends are getting the HPV vaccine”). A side-by-side comparison of the original CHIAS items and the modified items used for this analysis is described in We used all 16 items described in the original CHIAS today if it was available for you,” or “within the next 6 months” using a previously-published 11-point vaccination intention scale. Since you were in the lab for the first part of the study 6 months ago, have you received any doses (shots/injections) of the HPV vaccine?” (yes/no), and “Have you made an appointment to get the vaccine?” (yes/no).HPV vaccination intention and receipt were assessed as outcome variables. HPV vaccination intention was measured with two items that asked participants about the likelihood of getting the vaccine “An exploratory factor analysis of the baseline CHIAS items was conducted using principal components analysis with oblique rotation method (as factors were assumed to be correlated). Factors meeting the Kaiser criterion  were retained. Non-weighted factor scores (consistent with previous CHIAS assessments) p-values ≤0.05 were considered statistically significant. All analyses were performed using SPSS 20).Linear and logistic multivariable regression models were used to examine the association between the different factor groupings with vaccination intention and uptake, respectively. Each model included the 5 factors derived from the exploratory factor analysis, but no other covariates were added given our relatively small sample size (n = 139). For all analyses, Of the 139 participants who completed the baseline survey, 98 (70.5%) also completed the 6-month follow-up survey. As shown in When using a forced 4-factor solution, the exploratory factor analysis had good consistency in factor groupings with the original CHIAS for the “Barriers” and “Harms” factors. However, the statements loading to the “Effectiveness” and “Uncertainty” factors were markedly different from the original CHIAS under this solution strategy. When the analysis was repeated removing the restriction to 4 factors, the exploratory factor analyses demonstrated 5 factor groupings , which iincreased vaccination intention whereas increased concern about harms (Harms factor), lower perceived effectiveness (Effectiveness factor) and stronger risk denial attitudes  were associated with lower vaccination intention. Interestingly, when assessing vaccination intention for the coming 6 months, the Barriers and Risk Denial factors were no longer associated with this outcome (As shown in  outcome .Comparing Only 5 out of 98 women (5.1%) completing the 6-month follow-up assessment indicated that they had either received the HPV vaccine or made an appointment to get it since the baseline assessment. These low numbers precluded us from being able to perform any meaningful statistical analyses on how well the factors identified using CHIAS predicted vaccination uptake.Measures that reliably predict HPV vaccination intention across populations and over time could help facilitate the development of effective interventions to improve HPV vaccine uptake. The original CHIAS increased vaccination intention if the vaccine were available “today.” In contrast, at baseline there was no association between Barriers and vaccination intention when intentions for the “next 6 months” were assessed as the outcome, or when vaccination intention was assessed for either time frame in the follow-up survey. This finding could signify that the young women in the study had a very literal interpretation of having “the vaccine available for you today.” Participants may have believed that they would have opportunity to get the vaccine in the study lab after taking the baseline assessment (which was not the case). If so, it is understandable that those with higher perceived barriers to accessing the vaccine would have a higher vaccination intention for a vaccine that might be immediately available, and that access would be unrelated to a vaccine dose theoretically available 6 months in the future, or when reassessed by email where “vaccinating today” by the study team was obviously not a realistic possibility. These findings suggest that coupling HPV vaccination education with immediate access to the vaccine may be an effective strategy to increase HPV utilization among young adults.An interesting finding from our study was the association between the Barriers factor and vaccination intention. When assessed at baseline, young adults with higher perceived barriers to accessing the vaccine had an In the original CHIAS study in parents, CHIAS items appear to group into very similar factors when comparing parents making decisions about HPV vaccination for their adolescents to young women making the HPV vaccination decision for themselves, suggesting that the CHIAS is robust measure for categorizing HPV vaccination attitudes. However, the association of these factors with vaccination intention appears to differ between parents and young adults. Harms was the only factor that performed similarly between these two populations and also consistently predicted vaccination intention over a variety of time frames. This suggests that educational strategies focusing on mitigating perceived harms from the vaccine may have the widest influence and appeal across populations of different ages.


Detecting high-risk (HR) HPV is important for clinical management of women with persistent HPV-positive and Pap-negative results. The Cobas 4800 HPV test is the first FDA-approved HPV DNA test that can be used alone as a first-line screening tool. The HPV 9G DNA chip test is a PCR-based DNA microarray assay. We evaluated the patients of consecutive HPV-positivity on HPV 9G DNA chip test without cytologic abnormalities. We then compared the performances of HPV 9G DNA chip and the Cobas 4800 HPV tests for detecting HR HPV with each other and confirmed HPV genotyping using direct sequencing. All 214 liquid-based cytology specimens were collected from 100 women with consecutive HPV-positive and Pap-negative results on the HPV 9G DNA chip test between May 2012 and Dec 2013, but only 180 specimens were available for comparing HPV test results. The HPV 9G DNA chip and the Cobas 4800 HPV tests agreed with each other in 81.7% of the samples, and the concordance rate was greater than 97.2% for detecting HPV-16 or -18. For HR genotypes other than HPV types 16 and 18, the two tests agreed for 81.1% of the samples. The sensitivity of both assays for detecting HR HPV was 100%, regardless of HR genotypes. The HPV 9G DNA chip test may be as effective as the Cobas 4800 HPV test in detecting HR HPV, and has a similar ability to identify HPV-16 and -18. The Papanicolaou (Pap) screening test has significantly decreased the incidence of cervical cancer and its associated mortality during the last century. However, the Pap smear has low sensitivity and low reproducibility, in particular, for detecting high-grade squamous intraepithelial lesions (HSIL) . TherefoHPV infection, which is detected in 80% of SIL lesions, is the most common risk factor for cervical cancer . HPV genIn East Asia including Korea, the distribution of HPV genotypes is significantly different from that of Western populations. Specifically, HPV-58 is rare worldwide but is commonly found in East Asia –7. HPV-5Several HPV DNA genotyping tests (HPV DNA chips) are commercially available in Korea and have been widely used along with the Hybrid Capture assay because of their equivalent sensitivity and specificity for the detection of HSIL, as well as the concordance among various HPV DNA tests , 9. The In this study, we evaluated patients with consecutive HPV positivity without cytologic abnormalities using the HPV 9G DNA chip test and investigated the distribution and change in HPV genotypes. In addition, we compared the performances of the HPV 9G DNA chip and the Cobas 4800 HPV tests for detecting HR HPV DNA with each other and confirmed HPV genotyping using direct sequencing.This study was approved by the Institutional Review Board of the Catholic University of Korea, Incheon St. Mary’s Hospital (No. OC14EISI0081). No consent was given because data were analyzed anonymously. In total, 2023 women who were co-tested using liquid-based cytology and HPV 9G DNA chip test between May 2012 and December 2013 at Incheon St. Mary’s Hospital were selected after searching our electronic pathologic database. Of these 2023 women who underwent co-testing, 323 (16.0%) were determined to be cytology-negative but HPV-positive on the initial screening test without previous cytologic/histologic abnormalities; HR HPV , LR HPV , and HPV-other type . Among these 323 patients, 100 had two or more consecutive HPV-positive and Pap-negative results by successive HPV co-testing. Of these 100 patients, 14 patients had three consecutive HPV-positive and Pap-negative results. Finally, we collected all 214 cervical swab specimens from these 100 women who had consecutive HPV-positive and Pap-negative results for the study. All women did not undergo gynecologic procedures such as colposcopic biopsy, conization, or hysterectomy. All specimens were placed in a ThinPrep PreservCyt solution , and then stored at -70°C until they were assayed for HPV tests. The HPV 9G DNA chip and the Cobas 4800 HPV tests were performed by using the specimens from the same vials.HPV 9G DNA chip tests were performed and interpreted according to the manufacturer’s protocol . The whoCT from a sample is less than 40.0.HR HPV detection for the Cobas 4800 HPV test was also performed in all cervical swab specimens using the manufacturer’s protocol . The CobFor the samples with discrepant results between two HPV detection assays, their HPV genotypes were identified using PCR and sequencing. PCR was performed using the general primer MY09/11  and typek) with 95% confidence intervals (CIs), proportion of positive agreement (Ppos), proportion of negative agreement (Pneg), and McNemar’s p value were calculated to estimate the concordance between the results of two different tests. The k result was interpreted as follows: values < 0 as indicating no agreement and 0–0.20 as slight, 0.21–0.40 as fair, 0.41–0.60 as moderate, 0.61–0.80 as substantial, and 0.81–1 as almost perfect agreement. Ppos was calculated as twice the number of agreed positives/, while Pneg was calculated as twice the number of agreed negatives/. P values < 0.05 denoted statistical significance. Sensitivities, specificities, and 95% CIs of the Cobas 4800 HPV and the HPV 9G DNA chip tests were calculated based on the diagnostic accuracy criteria obtained from the genotyping results of the two HPV DNA tests and direct sequencing. When the results of two tests were concordant, they were regarded as true positive or negative. When there were discrepancies among the results, the diagnostic accuracy criteria were determined from the results of the HPV genotyping using direct sequencing.Statistical analyses were performed using Analyse—it Method Evaluation Edition, version 2.22, software . The agreement rate, kappa coefficient (n = 90) received short-term follow-up examinations for < 1 year.All 100 women with a median age of 49 years (range: 23–73 years) who were HPV-positive but Pap-negative at baseline were included in the analysis. Six women were under 30 years old. The follow-up interval between HPV co-testing ranged from 1.0 to 16.0 months . Most women . Of these 72 HR HPV-positive samples, 6 were co-infected with LR HPV.LR HPVs alone were seen in 20 samples (9.3%). HPV-other type was found in 122 samples (57.0%). HPV-16 was the most common genotype of HR HPV , either alone or in combination with other types. HPV-18 was observed alone in 7 samples. HPV-58 was the second most common genotype of HR HPV . All of HPV-58 presented as single infection, except for the one exhibiting co-infection with HPV-16. The remaining types of HR HPV included HPV-68 , HPV-66 , HPV-33 , HPV-52 , HPV-39 , HPV-51 , and HPV-56 . HPV-31, -35, -45, and -53 were infrequently observed (less than 5%).Only 180 out of 214 cervical swab specimens were available for comparison of HPV test results. Thirty four samples were not successfully analyzed by the Cobas 4800 HPV test due to a lack of remnant DNA or its degradation.The results of the Cobas 4800 HPV and HPV 9G DNA chip tests based on the detected genotypes of HR HPVs are summarized in k = 0.598) when detecting HR HPV , which was similar to the overall concordance rate between two assays regardless of HR HPV genotype (p < 0.05).When considering the HR genotypes other than HPV types 16 and 18, the results of the two HPV detection assays moderately agreed with each other at a rate of 81.1%  were negative for HR HPV genotypes, but the rest  showed different HR HPV genotypes on the Cobas 4800 HPV test. Among HR HPV-positive samples only on the Cobas 4800 HPV test, 20 were positive for HR genotypes other than HPV types 16 and 18; 11/20 samples were directly sequenced and no HR HPV was identified. One sample  was positive for HPV-16 on the Cobas 4800 HPV test, but no HR HPV was seen upon sequencing.LR HPVs were identified in 16 out of 180 specimens by the HPV 9G DNA chip test. Fourteen specimens  were negative for HR HPV genotypes on the Cobas 4800 HPV test. Only two samples (12.5%) were positive for HR genotypes other than HPV types 16 and 18, but none could be sequenced for genotyping.The sensitivities and specificities of the Cobas 4800 HPV and the HPV 9G DNA chip tests according to the detected genotypes of HR HPVs are summarized in HPV-positive/Pap-negative is the most common positive screening result obtained from co-testing. In a previous report in the Western population, 3.6% of women aged over 30 years have this co-test result at baseline . Song anSeveral previous reports have demonstrated the usefulness of HPV DNA chips in comparison with Hybrid Capture 2 in the general population –16. Diagp< 0.05). HPV-58 was the most frequently observed (n = 10) in the HR HPV genotypes other than HPV types 16 and 18, corresponding with previous studies [Nowadays, other HPV detection assays utilizing the real-time PCR method have been newly developed. The Cobas 4800 HPV test was recently accepted by FDA as a primary cervical cancer screening tool alone. The potential benefits of using the HPV DNA test for primary cancer screening are increasingly apparent . Howevern = 90) received short-term follow-up of HPV co-testing for < 1 year. Therefore, this might contribute to persistent HPV-positivity as well as various changes in HPV genotypes. Although the worldwide guidelines recommend retesting in 1 year for women with a baseline HPV-positive/Pap-negative screening results, such women return at varying intervals in practice [This study has a few limitations that should be discussed. First, most women in this study Click here for additional data file.