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副猪嗜血杆菌血清5型间接血凝试验和间接ELISA
抗体检测方法的建立
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石碧1，崔耀文1，贾凡1，金卉1，万云1，蔡旭旺1，方刚2，陈焕春1，周锐1
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（1.华中农业大学 动物医学院 农业微生物学国家重点实验室,湖北 武汉430070；
2.武汉水产科学研究所，湖北 武汉430065）
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摘要：
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为建立副猪嗜血杆菌（HPS）的血清学诊断方法，通过探索HPS荚膜多糖产生的最适体外培养条件，提取了HPS血清5型菌株的荚膜多糖（CPS），并以之为抗原分别建立了间接血凝试验（IHA）和间接ELISA两种抗体检测方法，对其特异性、敏感性和符合率进行了比较研究。结果表明，两种检测方法的特异性良好，但ELISA的敏感性是IHA的5～10倍，二者的阳性符合率、阴性符合率和总符合率分别为797%、55.2%和65.3%。用这两种方法检测了320份临床送检猪血清，IHA和ELISA的阳性率分别为40%和59%。结果证实，这两种方法适用于不同实验室条件下HPS的诊断和流行病学调查。 |
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关键词：
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副猪嗜血杆菌；荚膜多糖；间接血凝试验；间接ELISA |
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中图分类号：S 852.612：R 446.61 文献标识码：
A 文章编号：1673-4696(2007)11-0964-05 |
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Establishment of indirect haemagglutination and indirect ELISA assays
for detection of antibodies against Haemophilus parasuis serotype 5 |
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SHI Bi1,CUI Yao-wen1,JIA Fan1,JIN
Hui1,WAN Yun1,CAI Xu-wang1,
FANG Gang2,CHEN Huan-chun1,ZHOU Rui1 |
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(1.State Key Laboratory of Agricultural Microbiology/College of Veterinary Medicine,Huazhong Agricultural
University,Wuhan 430070,China;2.Wuhan Institute of Aquatic Science,Wuhan 430065,China) |
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Abstract: Indirect haemagglutination assay and ELISA were developed to detect antibodies against Haemophilus parasuis(HPS) serotype 5 in serum.Capsular polysaccharide (CPS) was pivotal in antigenicity and diagnosis HPS.The capacity of HPS serotype 5 to generate CPS in vitro was optimized.The CPS was purified from the HPS serotype 5 culture.IHA and ELISA for detection of the antibodies against HPS serotype 5 were developed using the purified CPS as antigen.The specificity,sensitivity and concordance of IHA and ELISA were compared.Both assays had high specificity; but the sensitivity of ELISA was approximately 5-10 times that of IHA.The concordance for the positive,negative and overall incidence between IHA and ELISA were 79.7%,55.2% and 65.3%,respectively.Of 320 porcine sera tested,made up 40% and 59% of samples showed positive results by IHA and ELISA,respectively.IHA and ELISA appeared to be two methods for diagnosis and epidemiological investigation of HPS under different laboratory conditions. |
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Key words: Haemophilus parasuis;capsular polysaccharide;IHA;indirect ELISA |
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