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通过用EcoRI进行DNA限制性内切酶分析来可靠鉴定单纯疱疹病毒。

Reliable identification of herpes simplex viruses by DNA restriction endonuclease analysis with EcoRI.

作者信息

Gerson M, Portnoy J, Hamelin C

出版信息

Sex Transm Dis. 1984 Apr-Jun;11(2):85-90. doi: 10.1097/00007435-198404000-00007.

Abstract

A simple procedure that permits the rapid identification of clinical isolates of herpes simplex virus (HSV, type 1 or type 2) on the basis of stable and type-specific differences in the EcoRI restriction endonuclease patterns of the HSV-1 and HSV-2 genomes was tested for reliability. Complete concordance was first obtained for 298 HSV isolates typed by standard laboratory procedures and by analysis of the DNA profiles. Two other HSV isolates showing intermediate biologic, immunologic, and/or biochemical properties were unambiguously identified by restriction endonuclease analysis of their genomes. Identical diagnostic results were also obtained by independent laboratories for another series of 150 HSV isolates after analysis of the EcoRI digestion products. As compared with other restriction enzymes, little intratypic variation in the DNA fragments selected for immediate identification of the isolates was found with EcoRI. The procedure used here, which does not require the purification and/or the radiolabeling of viral DNA, thus appears as most appropriate for the unambiguous typing of large numbers of HSV clinical isolates.

摘要

一种基于单纯疱疹病毒1型(HSV-1)和2型(HSV-2)基因组EcoRI限制性内切酶图谱的稳定且型特异性差异,快速鉴定临床分离的单纯疱疹病毒(HSV,1型或2型)的简单方法,对其可靠性进行了测试。通过标准实验室程序和DNA图谱分析对298株HSV分离株进行分型,首次获得了完全一致的结果。另外两株表现出中间生物学、免疫学和/或生化特性的HSV分离株,通过对其基因组进行限制性内切酶分析得到了明确鉴定。在对EcoRI消化产物进行分析后,独立实验室对另一组150株HSV分离株也获得了相同的诊断结果。与其他限制性酶相比,用EcoRI进行分离株直接鉴定时,所选DNA片段的型内变异很小。这里使用的方法不需要纯化和/或放射性标记病毒DNA,因此似乎最适合对大量HSV临床分离株进行明确分型。

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