Abstract: Objective Establishing the method of detecting the Japanese encephalitis virus(JEV) in swine and the biological materials from the swine with Immunofluorescence assay(IFA).Methods Selected JEV sensitive cell,titrate the virus’TCID50 and prepare the antigen slides according to GB IFA.titrate the best working density of immune fluorescence and carry out the experiments of specificity,sensitivity and stability.Result Selected BHK21 cell as JEV sensitive cell,the virus’ TCID50 is 10-8.9.mL-1;The best working density of immune fluorescence is 1∶ 200;There is no cross-reactivity with Classical swine fever virus(CSFV) and Porcine parvovirus(PPV),the experiments of sensitivity and stability show that the detection sensitivity is 1∶ 2 560 in different time;The lowest detected virus titer is 10-6.5.mL-1.Conclusion The IFA method is good in sensitivity,specificity and stability,as a result of which IFA can be used in detecting the swine and the biological materials from the swine.

Key words: Japanese encephalitis virus, Immunofluorescence assay

摘要： 目的建立乙型脑炎病毒(JEV)免疫荧光(IFA)检测方法,应用于猪及猪源性生物材料JEV的检测。方法滴定病毒TCID50,筛选JEV敏感细胞,依据国标IFA法制备抗原片,并进行特异性、敏感性和稳定性试验。结果选取BHK21细胞作为JEV敏感细胞,病毒感染力滴度(TCID50)为10-8.9.mL-1;与猪瘟病毒(CSFV)、猪细小病毒(PPV)均无交叉反应;稳定性和敏感性试验显示,不同时间IFA检测灵敏度均为1∶2560;可检测到的病毒滴度最低为10-6.5.mL-1。结论建立的IFA法敏感性、特异性强,稳定性好,可用于猪及猪源性生物材料JEV的检测。

关键词: 乙型脑炎病毒, 免疫荧光试验