检测狂犬病毒的胶体金免疫层析法的建立及应用

Gold-immunochromatography assay for detection of rabies antigens

目的 建立一种简易快速、适用于野外的胶体金免疫层析法 (GICA)用于检测动物脑或唾液中的狂犬病街毒抗原。方法 采用柠檬酸三钠还原法制备胶体金颗粒 ,标记抗狂犬病毒单抗 ,制成免疫层析测试条。样品中的狂犬病毒抗原与测试条上金标记抗体结合后沿着硝酸纤维素膜移动 ,与膜上的固相抗体结合形成肉眼可见的红色线条。结果 GICA测试条只与狂犬病毒抗原有特异性反应 ,与乙型脑炎病毒、正常小鼠脑组织等无交叉反应。GICA与EIA比较检测了 4 8份可疑狂犬病犬唾液样品 ,两法的符合率为 88 2 %。结论 GICA检测样品中的狂犬病毒抗原特异性强、、灵敏度高、简便快速、无需仪器设备 ,对基层单位开展初步鉴定狂犬病毒样品工作而言 ,不失为一种有效的方法。

To develop a sensitive,rapid and out-door used gold-immunochromotography assay (GICA) for detection of rabies antigens in the brain and saliva of suspicious rabid animals,colloidal gold was coupled with monoclonal antibodies aganst rabies to prepare the GICA apparatus.The gold-anti-rabies conjugate reacted with the rabies antigen in the brain or saliva of the suspicious rabid animals and migrated up to the nitrocellulose strip on which the anti-rabies antibody was immobilized,thus producing the detection reaction.It was found that the GICA testing strip only reacted with the rabies virus antigen to show a specific reaction,and showed no cross-reactions with Japanese B encephalitis virus antigen or normal mouse brain tissues.By using GICA and EIA in a comparative study of testing 84 specimens of saliva of suspected cases of patients with rabies,the corresphonding rate of both assays was 88.2%.It concludes that GICA is a simple and rapid method of assay for the detection of rabies antigens.