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副猪嗜血杆菌抗体间接血凝检测方法的建立及应用
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魏子贡1，蔡旭旺1，金梅林1,2，胡军勇1，葛君1，陈焕春1,2
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（1.华中农业大学 动物医学院 动物传染病室，湖北 武汉430070；
2.华中农业大学 农业微生物学国家重点实验室，湖北 武汉430070）
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摘要：
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将副猪嗜血杆菌（Haemophilus parasuis，HPS）血清型4、5型分离菌株经超声波破碎处理后的产物致敏醛化红细胞，建立了检测HPS抗体的间接血凝试验方法。最适反应条件为绵羊红细胞30℃醛化5h，4、5型菌株浓缩抗原以1∶8稀释致敏红细胞。免疫猪2周后检出率达89%。对15种HPS血清型阳性血清进行检测，结果均为阳性；对猪瘟病毒、口蹄疫病毒、猪圆环病毒、猪细小病毒、猪伪狂犬病病毒、猪生殖与呼吸综合征病毒、猪肺炎支原体、猪链球菌、猪肺疫巴氏杆菌、Ⅰ相支气管败血波氏杆菌及胸膜肺炎放线杆菌阳性血清进行检测，结果均为阴性。敏感性为琼脂扩散试验的16倍。对1665份猪血清进行检测，阳性率为47.1%。结果表明，该方法敏感性较高，特异性强，重复性好，可用于疫苗免疫后抗体水平的检测及副猪嗜血杆菌的流行病学调查。 |
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关键词：
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副猪嗜血杆菌；间接血凝试验；抗体检测 |
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中图分类号：S 852.612 文献标识码：
A 文章编号：1673-4696(2006)09-0713-06 |
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Establishment and application of an indirect hemagglutination assay for
detection of antibodies against Haemophilus parasuis |
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WEI Zi-gong1，CAI Xu-wang1，JIN
Mei-lin1,2，HU Jun-yong1，GE Jun1，CHEN
Huan-chun1,2 |
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(1.Laboratory of Animal Infectious Disease, College of Veterinary Medicine,Huazhong Agricultural
University, Wuhan 430070, China ; 2.National Key Laboratory of Agricultural Microbiology, Huazhong
Agricultural University,Wuhan 430070, China) |
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Abstract: An indirect hemagglutination assay(IHA) consisting of glutaraldehyde-fixed sheep red blood cells(SRBC) coated with the supernatant of sonicated HPS serovar 4 and 5 cells was established. SRBC could be best formalinized at 30℃ for 5 hours; The concentration of coating of antigen was 1∶8.The positive rate of vaccinated swine came to 89% two weeks later. All the fifteen positive sera of HPS showed positive results, and the positive sera of hog cholera virus, foot-and-mouth disease virus, porcine circovi-rus, porcine parvovirus, pseudorabies virus, porcine reproductive and respiratory syndrome virus, Mycoplasma hyopneumoniae, Streptococcus suis, Pasteurella multocida, Bordetella bronchiseptica and Actinobacillus pleuropneumoniae showed negative. The assay was 16 times more sensitive than agar-gel preci-pitation test. Examination of 1665 serum samples revealed that the positive ratio was 47.1%. These data indicated that the IHA established is a sensitive, specific, reproducible technique for detecting antibodies against Haemophilus parasuis. |
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Key words: Haemophilus parasuis; indirect hemagglutination assay(IHA); antibody detection |
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