微量板酶免疫斑点法测定麻疹病毒滴度  

Microplate Enzyme Immunospots (MEIS) for Titrating Measles Virus

作　　者：吕长国[1,2] 南一凡[1,2] 吴燕平[1,2] 贾芯 扈光伟 李守邦[1,2] 郑晓丽 

机构地区：[1]吉林省卫生防疫站 [2]卫生部长春生物制品研究所

出　　处：《中国计划免疫》1998年第2期63-66,共4页Chinese Journal of Vaccines and Immunization

摘　　要：测定麻疹病毒滴度常用微量板细胞病变法（ＣＰＥ）或蚀斑法。为简化实验结果判定过程，使实验指标判定更客观，我们在ＣＰＥ方法基础上建立了微量板酶免疫斑点法（Ｍｉｃｒｏｐｌａｔｅｅｎｚｙｍｅｉｍｍｕｎｏｓｐｏｔｓ，ＭＥＩＳ）。该方法通过酶促底物显色和抗原———抗体特异反应，使培养板上感染病毒造成细胞病变的部位形成可见的染色斑点。经肉眼直接观察，以培养板孔底出现特异性染色斑点为病变阳性，计算半数细胞感染浓度（ＣＣＩＤ５０），免去了显微镜下观察细胞病变的过程。经测定５０批麻疹病毒，ＭＥＩＳ法与ＣＰＥ法比较，结果ＣＣＩＤ５０符合率为９８．０％（４９／５０）；实验共用８００板孔，各孔结果总符合率为９９．８８％（７９９／８００），敏感性１００％（５１０／５１０），特异性９９．６６％（２８９／２９０）。本实验方法是敏感、特异的滴定麻疹病毒的新方法，尤其适用于多份病毒的滴定。An assay of microplate enzyme immunospots (MEIS) for titrating measles virus has been developed. The assay is based on cytopathic effect test (CPE) and employs technique of enzyme immune assays (EIA). The CPE produced by measles virus are presented as visible particular blue color spots in tissue culture microplate. In the MEIS, the cytopathogens are identified with naked eyes by means of finding particular color spots instead of observation under microscope for CPE. The MEIS was used in titrating 50 batches of measles virus and compared with CPE method. The CCID50 of these 50 batches were 98.0% (49/50) agreement with the results obtained from CPE method. The specificity and sensitivity was 99.66% and 100% respectively, calculated from the results of 800 wells in microplates. The assay is specific, sensitive, objective and applicable to titration of measles virus.

关 键 词：麻疹病毒 酶免疫分析 病毒滴度 CPE 麻疹疫苗 

分 类 号：R511.103[医药卫生—内科学]