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一种检测抗腮腺炎病毒中和抗体的新方法。

A new method for the detection of neutralizing antibodies against mumps virus.

机构信息

Department of Pediatrics, Hiroshima Prefectural Hospital, Minami-ku, Hiroshima City, Hiroshima, Japan.

出版信息

PLoS One. 2013 Jul 5;8(7):e65281. doi: 10.1371/journal.pone.0065281. Print 2013.

Abstract

Neutralization test is the most reliable method of evaluating immunity against viral diseases but there is no standard procedure for mumps virus, with tests differing in the infectivity of the challenge virus, 50% plaque reduction or complete inhibition of cytopathic effects (CPE), and usage of complement. A reliable, easy, and simple neutralization test for mumps virus was developed in this study. A recombinant mumps virus expressing GFP was generated as a challenge virus. Complement was added to the neutralizing mixture at 1∶200 when stocked serum samples were used. Neutralizing antibody titers were expressed as the reciprocal of the highest dilution that did not exceed two-fold of FU values (GFP expression) of the cell control wells. A total of 1,452 serum samples were assayed by inhibition of GFP expression in comparison with those examined by conventional 100% inhibition of CPE. 1,367 (94.1%) showed similar neutralizing antibody titers when examined by both methods. The GFP expression inhibition assay, using a recombinant mumps virus expressing GFP, is a simple and time- saving method.

摘要

中和试验是评估针对病毒性疾病免疫力最可靠的方法,但目前尚无腮腺炎病毒的标准程序,其检测方法在挑战病毒的感染力、50%蚀斑减少或完全抑制细胞病变效应(CPE)以及补体的使用上存在差异。本研究中开发了一种可靠、简便、易于操作的腮腺炎病毒中和试验。作为挑战病毒,构建了表达 GFP 的重组腮腺炎病毒。当使用储备血清样本时,在中和混合物中添加 1∶200 的补体。中和抗体滴度表示为细胞对照孔中 FU 值(GFP 表达)不超过两倍的最高稀释度的倒数。通过抑制 GFP 表达对 1,452 份血清样本进行了检测,并与通过传统方法检测的 100%抑制 CPE 进行了比较。两种方法检测的中和抗体滴度相似的有 1,367 份(94.1%)。使用表达 GFP 的重组腮腺炎病毒进行 GFP 表达抑制试验是一种简单、省时的方法。

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