Biased amplification of human papillomavirus DNA in specimens containing multiple human papillomavirus types by PCR with consensus primers.
Biased amplification of human papillomavirus DNA in specimens containing multiple human papillomavirus types by PCR with consensus primers.
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使用共有引物通过 PCR 对含有多种人乳头瘤病毒类型的样本中的人乳头瘤病毒 DNA 进行有偏差的扩增。
DOI：
10.1111/j.1349-7006.2011.01922.x
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
Kondo K. and Kanda T
中科院分区：
文献类型：
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作者：
Mori S.;Nakao S.;Kukimoto I.;Kusumoto-Matsuo R.;Kondo K. and Kanda T
临床标本中人乳头瘤病毒（HPV）的基因分型很重要，因为每种HPV类型具有不同的致癌潜力。根据HPV L1基因的保守序列设计引物，通过PCR扩增HPV DNA，已被广泛应用于基因分型。近年来研究表明，宫颈标本中往往含有多种类型的HPV，因此有必要确认使用共有引物的PCR是否能够无偏倚地扩增多种类型的HPV DNA。我们使用三种常用的共有引物对（L1 C1/L1 C2 +C2M，MY 09/11和GP 5 +/6+）通过PCR扩增测试样本中的HPV DNA，并通过与尼龙膜上的类型特异性探针杂交来鉴定所得扩增子。L1 C1/L1 C2 +C2M在含有HPV 16、18、51、52和58质粒之一的连续稀释的测试样品上显示出比其他引物更高的灵敏度，如通过检测HPV DNA的能力所定义的。L1 C1/L1 C2 +C2M不能扩增含有HPV 16、18或51的混合供试品中的HPV 16。这三种共有引物经常在含有HPV 16和HPV 18、31、51、52和58中的一种或两种的选定临床标本中引起不正确的基因分型。这些数据表明，PCR与共识引物是不适合的HPV基因分型标本中含有多种HPV类型，并建议通过这种方法获得的基因分型数据应仔细解释。（Cancer Sci2011; 102：1223-1227）
Genotyping human papillomavirus (HPV) in clinical specimens is important because each HPV type has different oncogenic potential. Amplification of HPV DNA by PCR with the consensus primers that are derived from the consensus sequences of theL1gene has been used widely for the genotyping. As recent studies have shown that the cervical specimens often contain HPV of multiple types, it is necessary to confirm whether the PCR with the consensus primers amplifies multiple types of HPV DNA without bias. We amplified HPV DNA in the test samples by PCR with three commonly used consensus primer pairs (L1C1/L1C2+C2M, MY09/11, and GP5+/6+), and the resultant amplicons were identified by hybridization with type‐specific probes on a nylon membrane. L1C1/L1C2+C2M showed a higher sensitivity than the other primers, as defined by the ability to detect HPV DNA, on test samples containing serially diluted one of HPV16, 18, 51, 52, and 58 plasmids. L1C1/L1C2+C2M failed to amplify HPV16 in the mixed test samples containing HPV16, and either 18 or 51. The three consensus primers frequently caused incorrect genotyping in the selected clinical specimens containing HPV16 and one or two of HPV18, 31, 51, 52, and 58. The data indicate that PCR with consensus primers is not suitable for genotyping HPV in specimens containing multiple HPV types, and suggest that the genotyping data obtained by such a method should be carefully interpreted. (Cancer Sci2011; 102: 1223–1227)