Genotyping and quantization of human papillomavirus by using the multi-fluorescent quantitative PCR assay
Objective Using the multi-fluorescent quantitative PCR assay to establish a fast-and-sensi- five screening method capable of quantization and genotyping of vires-load of human papillomaviruses(HPV)infec- tion for clinical diagnosis and therapy.Methods Two fluorescent dyes were used in a single PCR with type-spe- cific probes for HPV-16,HPV-33/52/58/67,HPV-18/45 and HPV-31.Quantization over a range of 5×10~1 to 5×10~7 copies/ml initial HPV copies is possible by using real-time detection of the accumulation of fluorescence with cycle number.A similar assay forβ-globing was used in parallei for absolute quantization of geuomic DNA. The above method is used on 120 samples and HC-2 has been used to calculate its specificity and sensitivity.Re- sults HPV DNA was detected in 63(52.5%)of 120 women cervical swabs by using the fluorescent quantitative PCR assay.HPV genotyping showed that HPV-16,HPV-18/45,HPV-31 and HPV-33/52/58/67 were detected in 36.51%(23/63),11.11%(7/63),12.70%(8/63),58.73%(37/63) of 63 samples,respectively.The average virus-load of HPV infection was 1.08×10~7 copies/ml.The specificity and sensitivity of this assay are 100% and 81.82%.Conclusion Our results demonstrates that the multi-fluorescent quantitative PCR assay is a simple, quick,sensitive and specific method for genotyping and quantization of HPV infection,it provides strong and de- tailed data for clinical screening on cervical specimens,virus-load on level of infection and objective method on re- suh of HPV infection therapy.