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ENZYME-LINKED IMMUNOSORBENT ASSAY(ELISA) FOR DETECTION OF MUMPS IgM ANTIBODY

酶联免疫吸附试验 (ELISA) 检测腮腺炎 IgM 抗体

基本信息

批准号：
60571079
负责人：
SAKATA HIROKO
金额：
$ 1.02万
依托单位：
NATIONAL INSTITUTE OF HEALTH
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60571079/
关键词：
Mumps IgM capture ELISA 血清学的診断

项目摘要

Serological disgnosis of viral infection is done by the demonstration of a significant increase of antibody during the period from acute to convalscent phases. Bleeding of patients at appropriate times is, however, not always possible and, in many instances, including mumps, only one serum specimen is available. Domonstration of IgM antibody provides the evidence for a recent infection with mumps virus. IgM antibody has usually been determined after separation of IgM fraction from a test serum by centrifugation through sucrose gradient, but the method is not easy for a routine serodiagnosis. We tried to develop a less sophisticated and less time-consuming procedure for determination of IgM antibody against mumps virus. We have developed the method in which IgM of a test serum is captured by anti-human IgM antibody attached to the solid phase. Purified mumps virus and peroxidase-conjugated monoclonal antibody specific to mumps virus NP protein then followed in succession. The color generated by the peroxidase in reaction with the substrate mixture was determined spectrophotometrically: In 24 cases of clinical mumps, the diagnosis of which had been confirmed by a significant rise in antibody titer in IgG-ELISA, IgM antibody was positive in 6 out of 9 sera taken on the first day of illness, in 8 out of 9 on day 2, and in all of 24 on day 5 and thereafter. A follow-up study with 25 cases showed that IgM antibody persisted for as long as 3 months and in one case up to 5 months. These results showed that the method is useful for the diagnosis of recent mumps virus infection.

病毒感染的血清学诊断是通过从急性期到恢复期抗体的显著增加来完成的。然而，在适当的时间出血的患者并不总是可能的，在许多情况下，包括腮腺炎，只有一个血清标本是可用的。IgM抗体的测定为近期流行性腮腺炎病毒感染提供了证据。IgM抗体通常是在通过蔗糖梯度离心从测试血清中分离IgM组分后测定的，但该方法对于常规血清学诊断来说并不容易。我们试图开发一种不太复杂和耗时较少的方法来测定腮腺炎病毒IgM抗体。我们开发了通过附着在固相上的抗人IgM抗体捕获测试血清中的IgM的方法。纯化的腮腺炎病毒和过氧化物酶标记的腮腺炎病毒NP蛋白特异性单克隆抗体，然后依次进行。过氧化物酶与底物混合物反应产生的颜色用荧光光度法测定：在24例临床腮腺炎病例中，其诊断已通过IgG-ELISA中抗体滴度的显著升高得到证实，在发病第一天采集的9份血清中有6份IgM抗体阳性，在第2天采集的9份血清中有8份IgM抗体阳性，在第5天及以后采集的所有24份血清中IgM抗体阳性。对25例患者的随访研究表明，IgM抗体持续时间长达3个月，1例长达5个月。结果表明，该方法可用于腮腺炎病毒近期感染的诊断。