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基于乳头瘤病毒氨基酸序列高度保守区域的简并引物可用于通用聚合酶链反应,以检测人乳头瘤病毒的有效感染。

Degenerate primers based on highly conserved regions of amino acid sequence in papillomaviruses can be used in a generalized polymerase chain reaction to detect productive human papillomavirus infection.

作者信息

Snijders P J, Meijer C J, Walboomers J M

机构信息

Department of Pathology, Free University Hospital, Amsterdam, The Netherlands.

出版信息

J Gen Virol. 1991 Nov;72 ( Pt 11):2781-6. doi: 10.1099/0022-1317-72-11-2781.

DOI:10.1099/0022-1317-72-11-2781
PMID:1658205
Abstract

Conserved amino acid sequences within the L1 open reading frame of the human papillomavirus (HPV) genome were used as a basis to design two degenerate primers (GP17 and GP18) and one general probe (GPR22) which direct polymerase chain reaction (PCR) amplification and subsequent detection of a 620 to 660 bp DNA fragment. The conserved nature of the primers and probe was tested experimentally on a panel of 24 cloned HPV DNAs isolated from cutaneous and mucosal lesions, including HPV-2a and -57, which are known to be associated with lesions at both anatomical sites. The sensitivity of this PCR test was at the level of genomic Southern blot analysis, indicating that HPV infections producing high copy numbers can be detected. Positive results were obtained with DNA extracted from clinical samples of genital and cutaneous origin.

摘要

人乳头瘤病毒(HPV)基因组L1开放阅读框内的保守氨基酸序列被用作设计两种简并引物(GP17和GP18)和一种通用探针(GPR22)的基础,这些引物和探针可指导聚合酶链反应(PCR)扩增并随后检测一个620至660 bp的DNA片段。在从皮肤和黏膜病变中分离出的24种克隆的HPV DNA组成的一组样本上,对引物和探针的保守性进行了实验测试,这些样本包括HPV - 2a和 - 57,已知它们与这两个解剖部位的病变相关。该PCR检测的灵敏度与基因组Southern印迹分析相当,表明可以检测到产生高拷贝数的HPV感染。从生殖器和皮肤来源的临床样本中提取的DNA获得了阳性结果。

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