间接ELISA检测鸭疫里氏杆菌抗体的研究

Development of an ELISA for the Detection of Antibody to Pasteurella Anatipestifer in Ducks

以提取的血清１型鸭疫里氏杆菌（Ｒ．ａ）Ｊｂ２株的脂多糖作为包被抗原，成功地建立了一种检测１型Ｒ．ａ抗体的间接ＥＬＩＳＡ方法，特异性试验和重复性试验证明此方法的特异性高、重复性好。并比较了间接ＥＬＩＳＡ方法、间接血凝试验、玻片凝集试验和琼脂扩散试验对抗体检出的敏感性，结果表明间接ＥＬＩＳＡ方法最敏感，人工感染后的鸭第７２小时即可检出血清抗体为阳性（３／４）；其次是间接血凝试验，第９６小时即可检出５０％（２／４）；玻片凝集试验在感染９天后才出现阳性（２／３）；而琼扩试验在感染后１１天仍为阴性反应。应用建立的间接ＥＬＩＳＡ方法，初步调查了南京地区鸭场此病的感染情况，发现所调查的１３日龄以前雏鸭的阳性率为０，而２１～３５日龄鸭的阳性率为８７３％。

An indirect ELISA was developed to detect specific antibody to Riemerella anatipestifer in sera of ducks.The Lipo polysaccharide antigen was used to coat ELISA plates,and assay was confirmed to be specific and reproducible. Experimentally infected ducks were assayed for specific antibody by the ELISA、indirect hemagglutination (IHA)、rapid slide agglutination (RSA) and Agar gel precipitation (AGP) test. Antibody to R.anatipestifer could be detected by the ELISA in some ducks at 3 days postinoculation (PI).Some sera were positive by the IHA at 4 days PI. Agglutinating antibody was detected by the RSA at 9 days PI. However,no precipitating antibody was detected by the AGP test in any of these samples at 11 days PI. Specific antibody to R.anatipestifer was detected in samples obtained from duck farms in Nanjing region by the ELISA. The data showed 6～13 day old ducklings had no specific antibody. However,87.3% of the investigated ducks which were 21～35 day old were positive.