用于测量单纯疱疹病毒和水痘-带状疱疹病毒对喷昔洛韦和阿昔洛韦敏感性的DNA探针检测法与蚀斑减少检测法的比较。

Comparison of a DNA probe assay with the plaque reduction assay for measuring the sensitivity of herpes simplex virus and varicella-zoster virus to penciclovir and acyclovir.

作者信息

Standring-Cox R, Bacon T H, Howard B A

机构信息

SmithKline Beecham Pharmaceuticals, Betchworth Surrey, UK.

出版信息

J Virol Methods. 1996 Jan;56(1):3-11. doi: 10.1016/0166-0934(95)01889-1.

DOI:10.1016/0166-0934(95)01889-1

PMID:8690764

Abstract

A DNA probe assay was compared with the plaque reduction assay to determine the sensitivity of clinical isolates of herpes simplex virus (HSV) and varicella-zoster virus (VZV) to penciclovir and acyclovir in MRC-5 cells. In both assays, penciclovir and acyclovir shared comparable activity against cell-free virus (CFV) preparations of VZV and herpes simplex virus type 1 (HSV-1) isolates, whilst acyclovir was significantly more active than penciclovir against herpes simplex virus type 2 (HSV-2) isolates in both the DNA probe assay (P < or = 0.01) and the plaque reduction assay (P < or = 0.01). However, the 50% effective concentrations (EC50s) were generally lower in the DNA probe assay and the correlation between the plaque reduction and DNA probe assays was poor for either compound. Six acyclovir-resistant strains of HSV-1 derived in cell culture were also tested for susceptibility to penciclovir and acyclovir, in the DNA probe and plaque reduction assays. The relative susceptibilities of these strains were comparable, for example, one ACV-resistant strain was susceptible to penciclovir in both assays. Further comparisons of the assay methods were made using cell-associated VZV (CAV). As with CFV the EC50s were significantly lower in the DNA probe assay than the plaque reduction assay for penciclovir (P < or = 0.01) and acyclovir (P < or = 0.01). In the DNA probe assay there was no significant difference in the EC50s for either penciclovir or acyclovir when comparing CAV with CFV. However, in the plaque reduction assay the EC50s for CAV were significantly higher than those for CFV for both penciclovir (P < or = 0.01) and acyclovir (P < or = 0.01). Overall the DNA probe assay is objective, does not require prior titration of isolates and provides opportunities for automation. It is more suitable for sensitivity testing of large numbers of clinical isolates than the well-established plaque reduction assay.

摘要

将DNA探针检测法与蚀斑减少检测法进行比较，以确定单纯疱疹病毒（HSV）和水痘带状疱疹病毒（VZV）临床分离株在MRC-5细胞中对喷昔洛韦和阿昔洛韦的敏感性。在这两种检测方法中，喷昔洛韦和阿昔洛韦对VZV和1型单纯疱疹病毒（HSV-1）分离株的无细胞病毒（CFV）制剂具有相当的活性，而在DNA探针检测法（P≤0.01）和蚀斑减少检测法（P≤0.01）中，阿昔洛韦对2型单纯疱疹病毒（HSV-2）分离株的活性均显著高于喷昔洛韦。然而，DNA探针检测法中的50%有效浓度（EC50）通常较低，且两种化合物的蚀斑减少检测法与DNA探针检测法之间的相关性较差。还在DNA探针检测法和蚀斑减少检测法中，对在细胞培养中获得的6株阿昔洛韦耐药HSV-1毒株进行了喷昔洛韦和阿昔洛韦敏感性测试。这些毒株的相对敏感性相当，例如，一株阿昔洛韦耐药毒株在两种检测方法中对喷昔洛韦均敏感。使用细胞相关VZV（CAV）对检测方法进行了进一步比较。与CFV一样，喷昔洛韦（P≤0.01）和阿昔洛韦（P≤0.01）的DNA探针检测法中的EC50显著低于蚀斑减少检测法。在DNA探针检测法中，比较CAV和CFV时喷昔洛韦或阿昔洛韦的EC50无显著差异。然而，在蚀斑减少检测法中，喷昔洛韦（P≤0.01）和阿昔洛韦（P≤0.01）的CAV的EC50均显著高于CFV。总体而言，DNA探针检测法客观，无需事先对分离株进行滴定，并为自动化提供了机会。与成熟的蚀斑减少检测法相比，它更适合对大量临床分离株进行敏感性测试。