Lei Yan-Jun, Gao Chen, An Run, Shi Qi, Chen Jian-Ming, Yuan Yu-Kang, Wang Chen, Han Jun, Dong Xiao-Ping
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Ying-Xin Road 100, Beijing 100052, China.
J Virol Methods. 2008 Jan;147(1):72-7. doi: 10.1016/j.jviromet.2007.08.005. Epub 2007 Sep 14.
The methods for detecting and typing human papillomavirus (HPV) in most molecular epidemiological surveys of verrucae vulgaris were based on PCR followed by sequencing or hybridization. However, the amplification efficacies of different assays for the detection of HPV DNAs varied largely. In this study, a novel multiplex PCR method to detect and type the HPVs (HPV-1, -2, -27 and -57) related to verrucae vulgaris was described. This method allows detecting and typing HPV DNA simultaneously in one reaction based on the length of the PCR products after electrophoresis. The sensitivity and specificity of this multiplex PCR method was assessed with the standard template panels and the spiking sample panels, and evaluated with the clinical samples, compared with PCR assay with primer MY09/11. The results showed the novel method had reliable clinical sensitivity (97.6%) and specificity (100%), significantly higher than that of the PCR using consensus primer, MY09/11. In addition, this method can effectively detect multiple HPV infection within the lesions. This simplified, economic and time-saving multiplex PCR method provides a useful additional tool for the clinical epidemiological study of verrucae vulgaris.
在大多数寻常疣分子流行病学调查中，检测和分型人乳头瘤病毒（HPV）的方法是基于聚合酶链反应（PCR），随后进行测序或杂交。然而，不同检测方法对HPV DNA的扩增效率差异很大。在本研究中，描述了一种用于检测和分型与寻常疣相关的HPV（HPV-1、-2、-27和-57）的新型多重PCR方法。该方法能够根据电泳后PCR产物的长度，在一个反应中同时检测和分型HPV DNA。使用标准模板板和加标样本板评估了这种多重PCR方法的敏感性和特异性，并与使用引物MY09/11的PCR检测法进行比较，用临床样本进行了评估。结果表明，该新方法具有可靠的临床敏感性（97.6%）和特异性（100%），显著高于使用通用引物MY09/11的PCR检测法。此外，该方法能够有效检测病变内的多种HPV感染。这种简化、经济且省时的多重PCR方法为寻常疣的临床流行病学研究提供了一种有用的补充工具。
