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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对11种人乳头瘤病毒进行基因分型

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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，快速且高通量的方法，用于同时检测11种人乳头瘤病毒（HPV）基因型，包括9种高风险类型（HPV16、18、31、33、35、39、52、58和66）和通过基于GenomeLab基因表达谱仪（GeXP）分析仪（GeXP-PCR）的多重PCR，可以在单个试管中使用两种低风险类型（HPV6和11）。使用11套嵌合引物启动PCR，并在随后的PCR循环中使用一对通用引物。用先前测试过的单一类型HPV感染的临床样品检查了GeXP-PCR对每种HPV类型的特异性。 GeXP-PCR的灵敏度通过对克隆的PCR产物进行连续10倍稀释的检测来评估。当含有HPV靶标的11种预混合质粒全部存在时，GeXP-PCR的灵敏度达到100拷贝。使用GeXP-PCR对124个临床标本进行的分析表明，GeXP-PCR测定法具有与报告的多重PCR测定法相当的敏感性和特异性，并且在混合感染的样本中HPV 11的检出率更高。总之，GeXP-PCR是检测多种HPV感染的快速，灵敏和高通量方法。

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