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原位杂交和免疫染色与细胞病理学在检测和鉴定培养细胞中单纯疱疹病毒感染方面的比较。

Comparison of in situ hybridization and immunologic staining with cytopathology for detection and identification of herpes simplex virus infection in cultured cells.

作者信息

Landry M L, Zibello T A, Hsiung G D

出版信息

J Clin Microbiol. 1986 Dec;24(6):968-71. doi: 10.1128/jcm.24.6.968-971.1986.

Abstract

Two recently developed sensitive techniques, in situ hybridization with a biotinylated cloned DNA probe and an avidin-biotin complex immunoperoxidase assay, were compared with the appearance of cytopathic changes for the early detection of herpes simplex virus infection in cell culture. By using commercially made reagents, these detection methods were evaluated in two different cell culture systems inoculated with both high- and low-input multiplicity of virus. The results revealed that both viral antigen and viral DNA detection methods could shorten the time to diagnosis of herpes simplex virus infection in cell culture; however, these methods were most useful in specimens containing low titers of virus when a less sensitive cell system was used. In this study, the avidin-biotin immunoperoxidase method was more sensitive and much cheaper than hybridization with a biotinylated probe. Significantly, when a highly sensitive cell system was used, cytopathic changes alone were comparable in rapidity and sensitivity to viral antigen or DNA detection methods applied in a less sensitive cell system.

摘要

将两种最近开发的灵敏技术,即使用生物素化克隆DNA探针的原位杂交技术和抗生物素蛋白-生物素复合物免疫过氧化物酶测定法,与细胞病变效应的出现情况进行比较,以用于在细胞培养中早期检测单纯疱疹病毒感染。通过使用市售试剂,在接种了高输入和低输入病毒感染复数的两种不同细胞培养系统中对这些检测方法进行了评估。结果显示,病毒抗原和病毒DNA检测方法均可以缩短细胞培养中单纯疱疹病毒感染的诊断时间;然而,当使用敏感性较低的细胞系统时,这些方法在含有低滴度病毒的标本中最为有用。在本研究中,抗生物素蛋白-生物素免疫过氧化物酶方法比用生物素化探针杂交更为灵敏且成本低得多。值得注意的是,当使用高敏感性细胞系统时,单独的细胞病变效应在快速性和敏感性方面与应用于敏感性较低细胞系统中的病毒抗原或DNA检测方法相当。

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