免疫印迹法在实验大鼠汉坦病毒监测中的应用及与间接免疫荧光法之比较

用汉坦病毒汉滩株（７６－１１８）重组核蛋白作为免疫印迹法（ＷｅｓｔｅｒｎＢｌｏｔ以下简称ＷＢ）的诊断抗原，用于实验感染大鼠血清抗体效价测定。同时与用汉城株（ＳＲ－１１）感染的Ｖｅｒｏ－Ｅ６细胞作抗原的间接免疫荧光法（以下简称ＩＦＡ）进行比较。ＷＢ法对３／４标本在大鼠接种病毒后第３天测得血清ＩｇＭ阳性，而ＩＦＡ法仅１／４标本出现阳性，ＩＦＡ效价为１：５１２０的血清，ＷＢ效价为１’：４０９６０，且在血清１：１０稀释时反应带亦清晰。两种方法分别测定６４份大鼠血清。甩ＩＦＡ法，４４份（６８．８％）出现类似阳性的荧光颗粒，而用ＷＢ法测定，无特异的反应带出现。非感染Ｖｅｒｏ－Ｅ６细胞作ＩＦＡ抗原，３０份（４６．９％）与正常细胞抗原有反应，此结果表明ＷＢ法在特异性和敏感性方面均高于ＩＦＡ法。ＩＦＡ法中的非特异性反应系血清与细胞成份之反应。

Western Blotting for Monitoring Hantavirus Antibodiesof Rats and Comparison with indirectImmunofluorescent Antibody

e developed Hantaanvirus ( 76-118) recombinant nucleocapsid protein( rNP ) as an antigen for Western Blotting ( WB ) to detect the antibodyVero E6 cell by Seoul virus (SR-11 ) as an antigen.Bb WB, 3 of 4 sera,butonly 1 of 4 by IFA, showed to be IgM antibody positive at the day 3 postinfection.The sera showed antibody titer of 1 :5120 by IFA and By WB.However, the sera showed a clear pattern even which was diluted from1 : 10 to 1 : 40969. The two methods were applied as the serologicalconfirmation of Hantavirus infection in 64 laboratory rats sera.By IFA,44 of 64 sera showed a strong IFA to infected Vero cell antigen, but, noneshowed a speciffc pattern by WB method.In addition, 30 of 64 sera alsoreacted with noninfected Vero cell antigen .The results demonstrated that thesensitivty and specificity of WB was suggested hiatsher than that of IFAtest. It was considered that the nonspecific immunofluorescence was theresponse to the cellular component.