GeXP多重PCR技术用于人乳头瘤病毒HPV检测和分型的研究

目的建立一种基于GenomeLab^TM GeXP（GenomeLab eXpress Profiling）遗传分析系统的人乳头瘤病毒（HPV）分型新技术。方法针对5种中国人常见的HPV亚型（HPV6,11,31,33和52）的基因序列进行比对分析,设计型特异性引物,建立并优化GeXP多重PCR反应体系,评价其特异性和灵敏性,并应用于30份临床标本检测。结果本研究建立的GeXP多重PCR技术能够成功检测和鉴别5种不同亚型的HPV病毒,灵敏度高,特异性好。结论成功建立了一种新的快速同时检测和鉴别5种HPV亚型病毒的多重PCR方法,适用于临床HPV感染的实验室诊断与流行病学研究。

Detection and typing of human papillomavirus by a GeXP based multiplex PCR assay

Objective To establish a new and rapid GeXP based multiplex PCR assay for the detection and typing of human papillomavirus 6, 11,31,33 and 52. Methods Nucleotide sequences of HPV6, HPV11, HPV31, HPV33 and HPV52 from NCBI were obtained and compared. Genotype-specific primers were then designed and the sensitivity and specificity of multiple PCR assay was evaluated.Optimized assay was further validated with 30 clinical specimens collected from the cervical secretions of patients. Results A GeXP based multiplex PCR was developed for sensitive detection and reliable differentiation of five HPV genotypes( HPV6,11,31,33 and 52) , Conclusion A GeXP based multiplex PCR assay is demonstrated to be a new and rapid technique for simultaneous detection and typing of 5 different human papillomaviruses.