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首页> 外文期刊>Journal of Medical Virology >Genotyping of 11 human papillomaviruses by multiplex PCR with a GeXP analyzer
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Genotyping of 11 human papillomaviruses by multiplex PCR with a GeXP analyzer

机译：用GEXP分析仪通过多重PCR进行11人乳头瘤病毒的基因分型

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摘要

A new, rapid, and high-throughput method was developed for simultaneous detection of 11 human papillomavirus (HPV) genotypes including nine high-risk types (HPV16, 18, 31, 33, 35, 39, 52, 58, and 66) and two low-risk types (HPV6 and 11) in a single tube by multiplex PCR based on a GenomeLab Gene Expression Profiler (GeXP) analyzer (GeXP-PCR). Eleven sets of chimeric primers were used to initiate the PCR, and one pair of universal primers was used for the subsequent cycles of the PCR. The specificity of GeXP-PCR for each HPV type was examined with clinical samples of single type HPV infection tested previously. The sensitivity of GeXP-PCR was evaluated by performing the assay on serial 10-fold dilutions of cloned PCR products. The GeXP-PCR achieved a sensitivity of 100 copies when all of the 11 pre-mixed plasmids containing HPV targets were present. Analyses of 124 clinical specimens using the GeXP-PCR demonstrated that the GeXP-PCR assay had comparable sensitivity and specificity to those of reported multiple PCR assay and an increased detection of HPV 11 in samples with mixed infections. In conclusion, the GeXP-PCR is a fast, sensitive, and high throughput method for the detection of multiple HPV infections.
机译：开发了一种新的，快速和高通量的方法，用于同时检测11人乳头瘤病毒（HPV）基因型（HPV）基因型，包括九种高风险类型（HPV16,18,31,33,35,39,52,58和66）和通过基于GenomeLab基因表达分析器（GEXP-PCR）的多重PCR，通过多重PCR在单管中（HPV6和11）中的两个低风险类型（HPV6和11）。使用11组嵌合引物引发PCR，并且一对环状引物用于PCR的后续循环。用先前测试的单型HPV感染的临床样品检查每种HPV型GEXP-PCR的特异性。通过在克隆的PCR产物的连续10倍稀释液上进行测定来评估GEXP-PCR的敏感性。当存在含有HPV靶标的11个预混合质粒时，GEXP-PCR实现了100份拷贝的敏感性。使用GEXP-PCR的124个临床标本分析表明，GEXP-PCR测定与报告的多PCR测定的那些具有相当的敏感性和特异性，以及在具有混合感染的样品中的样品中的HPV 11的检测增加。总之，GEXP-PCR是一种快速，敏感，高通量的检测多次HPV感染。

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