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用于扩增和检测生殖道人乳头瘤病毒类型的通用引物对。

A general primer pair for amplification and detection of genital human papillomavirus types.

作者信息

Evander M, Wadell G

机构信息

Department of Virology, University of Umeå, Sweden.

出版信息

J Virol Methods. 1991 Feb-Mar;31(2-3):239-50. doi: 10.1016/0166-0934(91)90162-s.

Abstract

A general primer pair localized in the E7 and E1 regions was identified and used for the detection of genital human papillomaviruses (HPVs). The genital HPV types 6b, 11, 16, 18, 31 and 33 were amplified and detected by the polymerase chain reaction (PCR) performed at a high stringency annealing temperature (60 degrees C). HPV-2, -3, -7, -13 and -30 were amplified only at lower temperatures. Twelve biopsies from women with invasive cancer in the cervix were analysed with the general primer pair. The amplification product specific for the general primer pair was detected in 11 of the 12 biopsies. The eleven HPV DNA positive specimens were shown to contain HPV-6b, HPV-16 and/or HPV-18 by Southern blot hybridization of the PCR products. The general primers were also used for analysis of 57 cervical scrapes from women with normal cytology, condyloma or CIN. By ethidium bromide staining after agarose gel electrophoresis we could detect 21 positives. Slot-blot analysis of the amplification products from all 57 scrapes confirmed the specificity of the 21 positives and revealed 5 additional positives. Among the 57 scrapes, 15/21 CIN scrapes, 10/21 condyloma scrapes and 1/15 normal scrapes contained HPV DNA. Eight different HPV types were detected. The general primer pair from the E7/E1 region is thus a powerful tool for the detection of HPV in clinical samples. The amplimer obtained offers a possibility for further typing by slot-blot hybridization using HPV-type specific probes.

摘要

鉴定出一对位于E7和E1区域的通用引物对,并将其用于检测生殖器人乳头瘤病毒(HPV)。通过在高严格度退火温度(60℃)下进行的聚合酶链反应(PCR)扩增并检测生殖器HPV 6b、11、16、18、31和33型。HPV - 2、- 3、- 7、- 13和- 30型仅在较低温度下扩增。用通用引物对分析了12例宫颈浸润癌女性的活检样本。在12例活检样本中的11例中检测到了通用引物对的特异性扩增产物。通过PCR产物的Southern印迹杂交显示,这11份HPV DNA阳性标本含有HPV - 6b、HPV - 16和/或HPV - 18。通用引物还用于分析57例细胞学正常、患有尖锐湿疣或宫颈上皮内瘤变(CIN)的女性的宫颈刮片。通过琼脂糖凝胶电泳后的溴化乙锭染色,我们检测到21例阳性。对所有57份刮片的扩增产物进行狭缝印迹分析,证实了21例阳性的特异性,并发现另外5例阳性。在57份刮片中,15/21例CIN刮片、10/21例尖锐湿疣刮片和1/15例正常刮片含有HPV DNA。检测到8种不同的HPV型别。因此,来自E7/E1区域的通用引物对是检测临床样本中HPV的有力工具。获得的扩增产物为使用HPV型特异性探针通过狭缝印迹杂交进行进一步分型提供了可能。

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