生殖器疣活检标本中HPV多重检测技术的建立

Establishments of multiple methods for the detection and typing of HPV in genital warts lesions

目的探索生殖器疣活检标本中人乳头瘤病毒(HPV)的多重检测分型技术,为我国生殖器疣感染HPV的检测和分型研究的开展提供有效手段。方法针对病毒L1区基因序列自行设计通用引物MY09、MY11、GP5+和GP6+,并对反应条件进行优化,建立基于聚合酶链式反应(PCR)及序列分析的检测生殖器疣活检标本中HPV感染型别的方法。结果采用巢氏PCR方法能够特异扩出140bp的目的条带,实现HPV感染的定性检测;通过对目的基因片段的测序分析能够实现单型别感染/多型别复合感染HPV的分型和鉴定,并可以检测到HPV少见的特殊类型,同时也能提供已知的HPV型别的突变信息。结论基于HPV病毒L1区基因序列建立的PCR和序列分析检测技术是快速检测和分型生殖器疣活检标本中HPV的一种有效方法。

Objective To develop an effective method for the detection of human papilloma virus （HPV） in genital warts lesions,and to provide standard techniques for the detection of HPV infection of genital warts in a multi-center clinical-epidemiology study. Methods Several pairs of oligo-nucleotide primers （MY09,MY11,GP5＋ and GP6＋） were designed and synthesized to amplify the specific DNA sequences by multiple PCR..Moreover,the reaction conditions of multiple PCR were optimized. Results The developed multiple PCR method could specifically amplify the expected sequences （140 bp）for HPV..Both the single-type and multiple-type HPV infection were successful detected,including the rare HPV strains and mutants. Conclusion A specific,sensitive,rapid and reliable PCR method was developed for the detection of HPV in genital warts lesions.