摘要
目的:摸索纯化麻疹病毒的方法,并以其为包被抗原,制备麻疹IgG酶联免疫(EIA)诊断试剂。方法:将L4株麻疹病毒液先后用超滤、PEG沉淀的方法进行浓缩,再分别用连续和不连续蔗糖密度梯度超速离心法进行纯化,比较不同方法纯化后的效果,最终建立制备高纯度麻疹病毒包被抗原的方法,并在此基础上制备麻疹IgG酶联免疫(EIA)诊断试剂。结果:用超滤、PEG沉淀及连续蔗糖密度梯度超速离心纯化后的麻疹病毒的比活性为775.76 HAU/mg,SDS-PAGE及Western-blot证明纯化产物的纯度符合要求,电镜结果显示纯化物中含大量麻疹病毒。以该纯化抗原制备的EIA检测试剂与血凝抑制实验(HI)相比,该试剂的灵敏度为99.46%,特异度为81.82%,一致性为97.58%。与SIGMA麻疹IgG EIA诊断试剂盒的一致率为95.12%。结论:采用超滤、PEG沉淀及连续蔗糖密度梯度超速离心的方法可制备满足麻疹IgG EIA诊断试剂要求的包被抗原。
Objective：To develop a method for preparation of coating antigen in measles IgG EIA test system. Methods：Measles virus IA strain tissue cultures was concentrated by ultrafiltration and PEG precipitation in order, the concentrated solution was then purified by continuous and discontinuous sucrose density gradient ultracentrifugation respectively. The purified results by deferent methods were compared and the final preparing methods established in purifying measles virus IA antigen. Results：The specific activities of IA measles virus purified by ultrafiltration, PEG precipitation and continuous sucrose density gradient ultracentrifugation was 775.76 HAU/mg. The purity of the purified virus was determined by SDS - PAGE and Western - blot. Typical measles virus particles were observed by transmission electron microscope in the purified viruses solution. As compared with hemagglutination -inhibitation（HI） test, the sensitivity of the kit which used purified IA measles virus as coating antigen to detect anti - measles IgG was 99. 46%, the specificity was 81.82%, and agreement was 97.58%. The agreement with SIGMA anti - measles IgG EIA kit was 95.12%. Conclusion：We can prepare the coating antigen in measles IgG EIA test system by continuous sucrose density gradient ultracentrifugation.
出处
《中国卫生检验杂志》
CAS
2007年第4期618-621,共4页
Chinese Journal of Health Laboratory Technology
