清洁级实验动物四种病毒抗体玻片酶免疫检测试剂盒的研制

本文报道对清洁级实验动物应排除的四种病毒（淋巴细胞脉络丛脑膜炎病毒、小鼠脱脚病病毒、鼠肝炎病毒和仙台病毒）抗体玻片酶免疫（ＥＩＡ）检测试剂盒的研制。四种病毒感染的细胞和对照细胞经冷丙酮固定于载玻片上制成特异性抗原和对照抗原，此四种病毒的抗血清各１０份和ＳＰＦ小鼠血清２０份分别与四种病毒的特异性抗原和对照抗原进行ＥＩＡ交叉试验，结果显示，抗原只与其相应抗血清发生特异性显色反应，与非特异性小鼠血清和ＳＰＦ小鼠血清不显色。与ＨＩ或ＥＬＩＳＡ方法比较，通过对１１２份普通小鼠血清进行测试，结果表明，ＥＩＡ对仙台病毒抗体的检出率（１９．６％）显著高于（＜０．００５）ＨＩ（６．３％），对小鼠脱脚病病毒抗体的检出率（２３．３％）与ＨＩ（２１．４％）无显著性差异（Ｐ＞０．０５）。ＥＩＡ对淋巴细胞脉络丛脑膜炎病毒和鼠肝炎病毒抗体的检出率分别为１．８％和７１．２％，ＥＬＩＳＡ对两种病毒抗体的检出率分别为１．８％和６７．６％，两种方法对两种病毒抗体的检出率无显著性差异（Ｐ＞０．０５）。重复性试验表明两批四种病毒抗体试剂盒对１０８份小鼠血清两次测定的符合率为９６～１００％。四种病毒的ＥＩＡ抗原在－１８℃保存１２个月或在２－８℃保存３

Studies on the Development of Slide EIA Kits for the Detection of Antibodies to Four Viruses That Should Be Removed from Clean Laboratory Animals

Studies on the development of slide enzyme immunoassay (EIA) kits for the detection of antibodies to four viruses (lymphocytic choriomeningitis virus, ectromelia virus, mouse hepatitis virus and Sendai virus) that should be removed from clean laboratory animals was reported.Culture cells infected respectively with these four viruses and uninfected cells were stabilized on the slides by acetone at 2-8℃ as specific and normal antigens. The cross EIA tests indicated that these virus antigens only reacted with their specific antisera respectively, but did not react with SPF (specific Pathogen free) mouse sera. Compared with HI or ELISA for detecting 112 conventional mouse sera, antibodies were detected to Sendai virus in 19.6% by EIA and in 6.3% by HI. There was prominant difference (P＜0.005) . Antibodies were detected to ectromelia virus in 23.3% by EIA and in 21.4% by HI. There was not prominant difference (P＞0.05) between them. Antibodies were detected to lymphocytic choriomeningitis virus in 1.8% and to moue hepatitis virus in 71.2% by EIA, and in 1.8% and in 67.6% by ELISA respectively. There was not prominant difference ( P＞0.05) between these two methods. 108 mouse sera were detected for antibodies to these four viruses by EIA with two lots of kits, indicating that the repetition rates were between 96-100%. After stored at -18℃ for twelvemonths or 2-8℃ for three months, the EIA antigens of these four viruses were still stable. In a word, the EIA kits for the detection of antibodies to these four viruses were cheap and easily performed as well as sensitive and specific.