小鼠抗体产生试验：我们能否不用它？

Mouse antibody production test: can we do without it?

作者信息

Mahabir E, Jacobsen K, Brielmeier M, Peters D, Needham J, Schmidt J

机构信息

Department of Comparative Medicine, GSF--National Research Centre for Environment and Health, D-85764 Neuherberg, Germany.

出版信息

J Virol Methods. 2004 Sep 15;120(2):239-45. doi: 10.1016/j.jviromet.2004.05.006.

DOI:10.1016/j.jviromet.2004.05.006

PMID:15288967

Abstract

Introduction of microbiologically contaminated materials into mice can cause infections of the recipients and jeopardize experimental protocols. As such, the methods used to screen biological materials should be sensitive, reliable and suitable for routine diagnostic work. In this report, the sensitivity of the viral plaque assay, mouse antibody production test and polymerase chain reaction (PCR) for detection of MHV-A59 and MMVp, two of the most prevalent pathogenic viruses in experimental mouse facilities, was compared. Analysis of serial tenfold dilutions of virus stocks revealed that the sensitivity of the mouse antibody production test on day 28 (10(-10) dilution) was at least 10 times higher than that of the viral plaque assay (10(-9) dilution) and 10(4) times more than that of the RT-PCR (10(-6) dilution) for detection of MHV-A59. For detection of MMVp, the PCR (10(-10) dilution) proved to be 10(6) times more sensitive than the viral plaque assay (10(-4) dilution) and the mouse antibody production test on day 28 (10(-4) dilution) which were equally sensitive. Based on the present study, it was shown that the method for diagnosis of viruses in biological materials should be employed only after the sensitivity has been determined for the viruses of interest implying that the most sensitive method needs to be determined independently for each virus.

摘要

将受微生物污染的材料引入小鼠体内会导致受体感染，并危及实验方案。因此，用于筛选生物材料的方法应灵敏、可靠且适用于常规诊断工作。在本报告中，比较了病毒蚀斑测定法、小鼠抗体产生试验和聚合酶链反应（PCR）对检测实验小鼠设施中两种最常见的致病病毒MHV - A59和MMVp的敏感性。对病毒原液的系列十倍稀释分析表明，在第28天进行的小鼠抗体产生试验（10⁻¹⁰稀释度）检测MHV - A59的敏感性比病毒蚀斑测定法（10⁻⁹稀释度）至少高10倍，比逆转录PCR（10⁻⁶稀释度）高10⁴倍。对于检测MMVp，PCR（10⁻¹⁰稀释度）的敏感性比病毒蚀斑测定法（10⁻⁴稀释度）高10⁶倍，而第28天的小鼠抗体产生试验（10⁻⁴稀释度）与之敏感性相同。基于本研究，结果表明只有在确定了针对目标病毒的敏感性之后，才应采用生物材料中病毒的诊断方法，这意味着每种病毒都需要独立确定最灵敏的方法。