Tucker R A, Johnson P R, Reeves W C, Icenogle J P
Viral Exanthems and Herpesvirus Branch, Centers for Disease Control and Prevention, Atlanta, GA 30333.
J Virol Methods. 1993 Aug;43(3):321-33. doi: 10.1016/0166-0934(93)90150-p.
Compared with other laboratory techniques, the polymerase chain reaction (PCR) is a simple, rapid, sensitive method for detecting human papillomavirus (HPV) DNA in cervical samples. However, since many cervical samples contain multiple HPV types, we decided to investigate whether PCR results from such samples accurately reflected the relative amounts of each HPV type present. Theoretical calculations of product accumulation when multiple DNAs with different amplification efficiencies are present in the same sample were done. In addition a set of samples in which cloned HPV DNAs were mixed in varying proportions prior to PCR was tested. Finally, four clinical samples containing multiple HPV types by hybridization assays were subjected to PCR, using two different primer sets. Each of these lines of investigation showed that selective amplification of one HPV DNA over another can occur when mixed HPV types are present. This effect may lead to inaccurate information regarding both types and relative amounts of HPV DNAs in samples containing multiple HPV types. A protocol to avoid this problem is described.
与其他实验室技术相比，聚合酶链反应（PCR）是一种用于检测宫颈样本中人乳头瘤病毒（HPV）DNA的简单、快速且灵敏的方法。然而，由于许多宫颈样本含有多种HPV类型，我们决定研究此类样本的PCR结果是否能准确反映每种HPV类型的相对含量。我们对同一样本中存在多种具有不同扩增效率的DNA时产物积累进行了理论计算。此外，还检测了一组在PCR之前将克隆的HPV DNA以不同比例混合的样本。最后，对四个经杂交检测含有多种HPV类型的临床样本，使用两种不同的引物组进行PCR。这些研究中的每一项都表明，当存在混合的HPV类型时，一种HPV DNA可能会比另一种发生选择性扩增。这种效应可能导致关于含有多种HPV类型样本中HPV DNA的类型和相对含量的信息不准确。本文描述了一种避免此问题的方案。
