乳胶凝集法快速检测猪圆环病毒2型  

Rapid Detection of Porcine Circovirus Type 2 by Latex Agglutination Test

作　　者：张慧敏[1] 杨利[1] 李广良[1] 王琳[1] 盛宗海[1] 韩鹤友[1] 

机构地区：[1]华中农业大学理学院,农业微生物国家重点实验室,武汉430070

出　　处：《分析化学》2011年第7期1113-1116,共4页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.20975042);华中农业大学自主科技创新重大培育专项基金(No.2010PY009);国家大学生创新实验计划(No.091050426)资助

摘　　要：发展了一种简单、快速、灵敏的乳胶凝集法用于猪圆环病毒2型的检测。合成了带有磺酸基的三元聚合物乳胶微球,通过静电相互作用,将猪圆环病毒2型单克隆抗体吸附到乳胶微球表面,形成乳胶抗体复合物诊断试剂。乳胶抗体复合物诊断试剂表面抗体与病毒表面抗原发生特异性反应,促使分散的乳胶微球凝集在一起,出现肉眼可见的凝集现象,而与去离子水、磷酸盐缓冲液、猪圆环病毒1型、猪繁殖与呼吸障碍综合症病毒、猪细小病毒、猪传染性胃肠炎病毒不出现凝集现象。在最优化条件下,病毒浓度检测范围为3.1×105～8.00×107 copy/mL,检出限为1.0×105 copy/mL。采用本方法与PCR方法检测了34份临床血清样本,结果的符合率达到94.1%,敏感性为89.5%,特异性达到了100%。结果表明,本方法可用于临床血清样品的检测,在猪圆环病毒2型的检测中表现出良好的应用前景。A facile,rapid,ultrasensitive and visual method was developed for the determination of porcine circovirus type 2（PCV2） based on latex agglutination test.Ternary polymer latex particles with sulfonic acid group were synthesized,then the monoclonal antibody of PCV2 bound to latex particles by electrostatic interactions and generated latex-antibody compound.The specific reaction of monoclonal antibody and virus showed visible agglutination phenomenon in the presence of a certain concentration of virus.Latex-antibody compound didn′t react with deionized water,phosphoric acid buffer,porcine circovirus type 1,porcine reproductive and respiratory syndrome virus,porcine parvovirus and porcine transmissible gastroenteritis virus.Under the optimized experimental conditions,the detection range of PCV2 was from 3.1×105 copy/mL to 8.0×107 copy/mL,and the detection limit was 1.0×105 copy/mL.Compared with PCR,the proposed method has good efficiency（94.1%）,sensitivity（89.5%） and specificity（100%） in 34 serum samples analysis.

关 键 词：猪圆环病毒2型 单克隆抗体 乳胶凝集 

分 类 号：S858.28[农业科学—临床兽医学]