Studies On Establishment Of Detection Method For HPV Infection Using Fluorescence Quantitative PCR

Objective: The research was to discuss the feasibility of fluorescence quantitative PCR (FQ-PCR) in screening the infection of Human paplimavirus in feminine, and to explore an experimental basis and theoretical foundation for the clinical application.Methods:①Collected 600 cases of uteri cervix secretion, extracted their DNA followed by the instruction of kits, and detected HPV DNA copies by FQ-PCR technology. Regarded specimens whose DNA copies were higher than the lowest detection limit were suspicious specimens;②Analysed HPV subtypes which suspicious specimens may probably be infected with by use of the technology of membrane strip hybrid;③Verified the accuracy of FQ-PCR results by using DNA sequence determination technology.Results:①A total of 63 cases were suspected to be infected with HPV. The results of fluorescence quantitative PCR showed that the initial content of HPV were higher than 50 copies/ml, and the specimens were regarded as suspicious positive samples. The positive rate was 10.5%. The group ,≤ 35-year-old , got the highest positive rate, followed by 35 to 50-year-old group and≥50-year-old group.②In the experiment of membrane hybridization , there were 62 cases of suspicious positive samples which appeared specific blue dots, with the positive rate of 98.41%. We also found that the main HPV subtypes spicimens infected were HPV16, HPV 58, HPV 18. And most of the infection were only detected by a single-type HPV.③The sequencing results, which sample was infected with a single type of HPV, had the same conservative region with HPV L1 gene sequence that Genebank provided.Conclusion: Both the results of hybridization and sequencing confirmed the accuracy of fluorescence quantitative PCR on testing HPV infection. The results got a high credibility. Additonally, because of its advantadges as high-speed, full-closed response, high-throughput advantages, it can be considered that it was able to be used for testing female HPV infection in population census. And it was meaningful to prevent cervical cancer in combination with other auxiliary inspections.