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用于检测人乳头瘤病毒DNA的Southern印迹杂交法与聚合酶链反应法的比较

Comparison of Southern blot hybridization and polymerase chain reaction methods for the detection of human papillomavirus DNA.

作者信息

Schiffman M H, Bauer H M, Lorincz A T, Manos M M, Byrne J C, Glass A G, Cadell D M, Howley P M

机构信息

Environmental Epidemiology Branch, National Cancer Institute, Bethesda, Maryland 20892.

出版信息

J Clin Microbiol. 1991 Mar;29(3):573-7. doi: 10.1128/jcm.29.3.573-577.1991.

Abstract

A methodologic study was performed to compare the polymerase chain reaction (PCR) and Southern blot hybridization, two commonly used testing strategies for the detection of human papillomavirus (HPV) infection. Three laboratories tested masked aliquots of exfoliated cervical cell specimens obtained from 120 women by cervicovaginal lavage. The study population included 32 women with condylomatous atypia or cervical intraepithelial neoplasia and 88 control women with no known history of cervical neoplasia. Two laboratories used PCR with different sets of consensus primers for HPV detection. The third laboratory used low-stringency Southern blot hybridization to identify all HPV types, followed by high-stringency Southern and/or dot blot hybridization to confirm specific HPV types. One of the PCR primer sets detected HPV types with a differential efficiency that was not predicted by analysis of DNA sequences or direct testing of HPV-containing plasmids. In contrast, the second PCR primer set was shown to be a much broader consensus system, detecting the same HPV types as Southern blotting, though requiring much less clinical specimen. Over 80% of women with cervical intraepithelial neoplasia or condylomatous atypia were found to be HPV infected both by Southern blotting and by the second PCR primer set. Among the control women, 11% were HPV positive by Southern blotting, while 31% were positive with the second set of primers. Most of the HPV infections found only by PCR were not due to HPV type 6, 11, 16, 18, 31, 33, or 45. These known HPV types were uncommon among normal women in the study population, even as determined by the PCR method.

摘要

开展了一项方法学研究,以比较聚合酶链反应(PCR)和Southern印迹杂交这两种常用于检测人乳头瘤病毒(HPV)感染的检测策略。三个实验室对通过宫颈阴道灌洗从120名女性获取的脱落宫颈细胞标本的盲法等分试样进行检测。研究人群包括32名患有湿疣样异型或宫颈上皮内瘤变的女性以及88名无宫颈瘤变病史的对照女性。两个实验室使用不同组的HPV检测通用引物进行PCR检测。第三个实验室使用低严谨度的Southern印迹杂交来鉴定所有HPV类型,随后使用高严谨度的Southern和/或斑点印迹杂交来确认特定的HPV类型。其中一组PCR引物检测HPV类型的效率存在差异,这在对DNA序列的分析或对含HPV质粒的直接检测中并未预测到。相比之下,第二组PCR引物显示为一个更具通用性的系统,能检测到与Southern印迹杂交相同的HPV类型,不过所需临床标本要少得多。通过Southern印迹杂交和第二组PCR引物检测发现,超过80%患有宫颈上皮内瘤变或湿疣样异型的女性感染了HPV。在对照女性中,Southern印迹杂交显示11%的女性HPV呈阳性,而第二组引物检测显示31%的女性呈阳性。仅通过PCR检测到的大多数HPV感染并非由6、11、16、18、31、33或45型HPV所致。这些已知的HPV类型在该研究人群的正常女性中并不常见,即便通过PCR方法检测也是如此。

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