肺炎球菌免疫血清库的建立及应用  

Set up a pneumococcal antisera bank and related application

作　　者：刘倩 张丽芝 张新庄 任克明 陈晓航 王剑虹 张钦 沈荣 LIU Qian ZHANG Li-zhi ZHANG Xin-zhuang REN Ke-ming CHEN Xiao-hang WANG Jian-hong ZHANG Qin SHEN Rong(First Department of Research, Lanzhou Institute of Biological Products Co. , Ltd. , Center for Gansu Provincial Vaccinc Engineering Research, Lanzhou 730046, Gansu Province, Chin)

机构地区：[1]兰州生物制品研究所有限责任公司第一研究室甘肃省疫苗工程技术研究中心,甘肃兰州730046

出　　处：《微生物学免疫学进展》2017年第5期1-10,共10页Progress In Microbiology and Immunology

基　　金：国家高技术研究发展计划项目(2012AA02A401)

摘　　要：目的建立24个血清型的肺炎链球菌(简称肺炎球菌)免疫血清库,为肺炎球菌疫苗中各型多糖含量准确测定提供可靠保障。方法 (1)制备肺炎球菌各型免疫血清:用肺炎球菌菌株制备免疫原,按免疫程序接种青紫蓝兔,将符合规定的免疫血清合并、除菌、分装后建档;(2)鉴定肺炎球菌各型免疫血清:全程采用速率散射比浊法进行鉴定。首先确定肺炎球菌免疫血清工作浓度,再通过特异性和线性对免疫血清进行初步筛查(将筛查出有交叉反应的免疫血清进行吸收),然后用血清交叉试验进行再次筛查,最后通过绘制反应曲线(截取多糖质量浓度1.0~5.0μg/m L的线段)对免疫血清进行线性评定;(3)肺炎球菌免疫血清库的应用:用建成的肺炎球菌免疫血清库中各型免疫血清和丹麦国家血清研究所的诊断血清,检测5个厂家的23价肺炎球菌多糖疫苗的各型多糖含量,对比结果。结果建成的肺炎球菌免疫血清库各型免疫血清效价较高;初步筛查出的有交叉反应的免疫血清,经吸收后交叉反应消失且线性良好(R2>0.98);通过血清交叉试验再次筛查,未见有交叉反应的免疫血清;反应曲线线段(多糖质量浓度1.0~5.0μg/m L)线性良好(R2>0.98);应用两套血清检测23价肺炎球菌多糖疫苗的多糖含量,差异无统计学意义(P>0.05)。结论成功建立了肺炎球菌免疫血清库,且各型免疫血清稳定、无交叉反应,可用于肺炎球菌疫苗及其疫苗血清型多糖的各项免疫学检测。Objective A pneumococcal antisera bank contained 24 serotypes of antiserums was set up,in order to provide a reliable guarantee in accurate determination of related polysaccharides contents in streptococcus pneumoniae vaccine.Methods（1） All types of pneumococcal antiserums were prepared by using each serotype of pneumococcal strains as immunogen,respectively,the Chinchilla rabbits were immunized according to immunization schedule. The antiserums were mixed,filter-sterilized,aliquoted certified,then filing;（2） The identification was carried out by rate nephelometry. First,the working concentration of pneumococcal antisera was determined. Secondly preliminary screening of antiserums were performed in both tests for specificity and linearity（ the screened antiserums with cross-reaction were absorbed）. Thirdly,all antiserums were performed by serocross test. Finally,each type of antiserum was evaluated linearity by plotting the reaction curve（ intercept the line segment of 1. 0-5. 0 μg/m L of polysaccharide mass concentration）;（3） Application： 23 valent pneumococcal polysaccharide vaccine from 5 manufacturers were tested by the pneumococcal antisera banks,which included the set bank and the bank containing diagnostic serums form Statens serum institut,and the results were compared. Results The each type serum contained in pneumococcal antisera bank has a higher titer,and the cross-reaction was removed and showed a good linearity after absorption（ R^2〉0.98）. There was no cross-reaction in all antiserums screened by serocross test. The response curve segment（ the line segment of 1.0-5.0 μg/m L of polysaccharide mass concentration） showed a good linearity（ R^2〉0.98）,by using the two banks to detect the polysaccharide content in the vaccine,and tested difference had no a statistical meaning（ P〈0.05）. Conclusion The pneumococcal antisera bank was successfully prepared,and all of contained antiserums were stable and no cross-reaction,it can be used in immunological detection of

关 键 词：肺炎球菌免疫血清 血清型 多糖 血清库 速率散射比浊法 

分 类 号：R392-33[医药卫生—免疫学]