Highly effective detection of human papillomavirus 16 and 18 DNA by a testing algorithm combining broad-spectrum and type-specific PCR
Highly effective detection of human papillomavirus 16 and 18 DNA by a testing algorithm combining broad-spectrum and type-specific PCR
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DOI：
10.1128/jcm.00539-06
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发表时间：
2006-09-01
影响因子：
9.4
通讯作者：
Colau, Brigitte
中科院分区：
文献类型：
--
作者：
van Doorn, Leen-Jan;Molijn, Anco;Colau, Brigitte
由于混合感染中不同基因型之间的竞争，使用单一的基于广谱人乳头瘤病毒（HPV）DNA的PCR检测可能无法检测出较低浓度的HPV DNA。为了提高PCR检测HPV的能力，将广谱和类型特异性（TS）PCR相结合，重点关注HPV-16和HPV-18。从参与HPV-16/HPV-18候选疫苗有效性试验的1，113名健康女性（年龄范围，15至25岁）中获得宫颈和宫颈阴道细胞样本。这些样本通过广谱SPF 10 PCR-DNA酶免疫测定法进行检测，然后通过基于引物SPF 10的线性探针测定法（SPF 10利帕）以及HPV-16和HPV-18-TS PCR进行检测。大多数HPV-16/18 SPF 10 LiPA阳性样本的结果通过TS-PCR得到证实（HPV-16的kappa值为0.775，HPV-18的kappa值为0.785）。然而，TS PCR显示，由于竞争，在含有其他HPV基因型的样本中有额外的阳性样本。相反，SPF 10利帕在由于采样变异而通过TS PCR保持阴性的样本中鉴定出HPV-16或-18。对1,000多名妇女的随访样本的分析证实，SPF 10-利帕与额外的HPV-16-和HPV-18-TS PCR的组合降低了假阴性诊断率。广谱PCR和TS PCR的结合产生了一种新的检测算法。这种检测方法的组合比单独使用任何一种方法都更准确，新算法为HPV感染的分析提供了高度准确和有效的方法。
The use of a single broad-spectrum human papillomavirus (HPV) DNA-based PCR test may fail to detect lower concentrations of HPV DNA due to competition between different genotypes in mixed infections. To improve HPV detection by PCR, broad-spectrum and type-specific (TS) PCRs were combined, with a focus on HPV-16 and HPV-18. Cervical and cervicovaginal cell samples were obtained from 1,113 healthy women (age range, 15 to 25 years) participating in an HPV-16/HPV-18 candidate vaccine efficacy trial. These samples were tested by a broad-spectrum SPF10 PCR-DNA enzyme immunoassay, followed by a primer SPF10-based line probe assay (SPF10 LiPA), and HPV-16- and HPV-18-TS PCRs. The results for the majority of the HPV-16/18 SPF10 LiPA-positive samples were confirmed by TS-PCR (kappa values, 0.775 for HPV-16 and 0.785 for HPV-18). However, TS PCR revealed additional positive samples among those that contained other HPV genotypes due to competition. Conversely, SPF10 LiPA identified HPV-16 or -18 in samples that remained negative by TS PCR as a result of sampling variation. Analysis of follow-up samples from more than 1,000 women confirmed that the combination of SPF10-LiPA with additional HPV-16- and HPV-18-TS PCR diminishes the rate of false-negative diagnosis. The combination of broad-spectrum and TS PCRs resulted in a novel testing algorithm. This combination of assays is more accurate than either method alone, and the novel algorithm offers a highly accurate and effective method for the analysis of HPV infections.