论文部分内容阅读
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。
Measles measles virus titer commonly used microplate cytopathic method (CPE) or plaque. In order to simplify the determination process of experimental results and make the determination of experimental indexes more objective, we established Microplateenzymeimmunospots (MEIS) based on the CPE method. The method of enzymatic substrate chromogenic and antigen - antibody-specific reaction, so that the culture plate infected with virus causes cytopathic site formation of visible staining spots. Direct observation with the naked eye, the bottom of the hole in the culture plate appeared stain positive for the lesions, the median concentration calculated (CCID50), eliminating the need for microscopic observation of cytopathic process. After 50 batches of measles virus were determined, the coincidence rate of MEIS and CPE was 98.0% (49/50). The experiment shared 800 wells, the total coincidence rate of each well was 99.88% (799/800 ), The sensitivity was 100% (510/510) and the specificity was 99.66% (289/290). The experimental method is sensitive and specific titration of measles virus new method, especially for multiple titration of the virus.