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

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79.7%、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。

Establishment of indirect haemagglutination and indirect ELISA assays for detection of antibodies against Haemophilus parasuis serotype 5

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.