Oevestad Irene T, Janssen Emiel A M, Baak Jan P A
Department of Pathology, Stavanger University Hospital, Stavanger, Norway.
Indian J Pathol Microbiol. 2007 Oct;50(4):733-9.
High-risk human papillomavirus (hr-HPV) testing is widely used, but different DNA isolation methods may give variable results in PCR determinations. To evaluate the effect of different DNA isolation methods on the HPV results, DNA was isolated from 180 consecutive cervical samples with the manual Amplilute kit, or the standard automated or a modified MagNAPure LC (MPLC) protocol. The DNA was then tested for hr-HPV with the AMPLICOR HPV-test, and the HPV-genotype was determined with the Linear Array (LA) test. The concordance of the three DNA isolation methods is 90-93% and 88-92% between the HPV-tests from DNA isolated by any of the three isolation methods and LA. The sensitivity, specificity, percentage of false negatives and false positives compared to the LA detected hrHPV subtypes are similar for the manual and standard MPLC, but the specificity of the modified MPLC was much lower and the percentage of false positives much higher. In conclusion both the manual and standard MPLC DNA isolation methods give good HPV results. The standard MPLC is the fastest DNA isolation method.
高危型人乳头瘤病毒(hr-HPV)检测被广泛应用,但不同的DNA提取方法在PCR检测中可能会得出不同的结果。为评估不同DNA提取方法对HPV检测结果的影响,我们使用手动Amplilute试剂盒、标准自动化方法或改良的MagNAPure LC(MPLC)方法,从180份连续的宫颈样本中提取DNA。然后使用AMPLICOR HPV检测试剂盒检测提取的DNA中的hr-HPV,并通过线性阵列(LA)检测确定HPV基因型。三种DNA提取方法之间的一致性在90%-93%之间,且通过这三种提取方法中的任何一种提取的DNA进行HPV检测所得结果与LA检测结果之间的一致性在88%-92%之间。与LA检测出的hrHPV亚型相比,手动提取方法和标准MPLC方法的灵敏度、特异性、假阴性和假阳性百分比相似,但改良MPLC方法的特异性要低得多,假阳性百分比要高得多。总之,手动提取方法和标准MPLC DNA提取方法都能得出良好的HPV检测结果。标准MPLC方法是最快的DNA提取方法。