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通过非放射性自旋扩增原位杂交试验检测单纯疱疹病毒

Detection of herpes simplex virus by a nonradiometric spin-amplified in situ hybridization assay.

作者信息

Forman M S, Merz C S, Charache P

机构信息

Department of Laboratory Medicine, Johns Hopkins Medical Institutions, Baltimore, Maryland 21205.

出版信息

J Clin Microbiol. 1992 Mar;30(3):581-4. doi: 10.1128/jcm.30.3.581-584.1992.

Abstract

An in situ hybridization kit (Diagnostic Hybrids, Inc., Athens, Ohio) was evaluated for use in the detection and identification of herpes simplex virus (HSV) from clinical specimens. For in situ hybridization, a 10-min spin amplification onto monolayers of African green monkey kidney cells (CV-1) in 24-well polystyrene dishes, 24-h culture amplification, and hybridization with an alkaline phosphatase-labeled DNA probe were used. A total of 648 specimens were tested, including 275 specimens from patients with symptomatic diseases sent specifically for HSV detection and 373 specimens from asymptomatic immunocompromised patients sent for detection of HSV shedding. Overall, the sensitivity of the hybridization assay was 97.8% (131 of 134 specimens), with 105 of 105 (100%) specimens from symptomatic patients and 26 of 29 (89.9%) specimens from asymptomatic patients being detected. The three specimens that were false negative by in situ hybridization had low virus titers, as determined by tissue culture. The specificity was 99.6% (512 of 514 specimens). The rapid, accurate results suggest that the in situ hybridization kit may be used as an alternative to conventional tissue culture for the detection of HSV.

摘要

对一种原位杂交试剂盒(Diagnostic Hybrids公司,俄亥俄州雅典市)进行了评估,以用于从临床标本中检测和鉴定单纯疱疹病毒(HSV)。对于原位杂交,采用了在24孔聚苯乙烯培养皿中的非洲绿猴肾细胞(CV-1)单层上进行10分钟的旋转扩增、24小时的培养扩增以及与碱性磷酸酶标记的DNA探针杂交的方法。总共检测了648份标本,包括专门送检用于HSV检测的275份有症状疾病患者的标本以及送检用于检测HSV脱落的373份无症状免疫功能低下患者的标本。总体而言,杂交检测的敏感性为97.8%(134份标本中的131份),其中有症状患者的105份标本(100%)和无症状患者的29份标本中的26份(89.9%)被检测到。通过原位杂交呈假阴性的三份标本,经组织培养测定病毒滴度较低。特异性为99.6%(514份标本中的512份)。快速、准确的结果表明,原位杂交试剂盒可作为检测HSV的传统组织培养方法的替代方法。

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