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当前位置: 首页 > 文章 > 猪嗜血支原体酶标单抗阻断ELISA检测方法的建立 畜牧兽医学报 2011,42 (11) 1591-1597
Position: Home > Articles > Development of Blocking ELISA for Mycoplasma suis with Peroxidase-Labelled Monoclonal Antibody Against MSG1 Protein Chinese Journal of Animal and Veterinary Sciences 2011,42 (11) 1591-1597

猪嗜血支原体酶标单抗阻断ELISA检测方法的建立

作  者：
张长莹;李玉峰;姜平
单  位：
南京农业大学动物疫病诊断与免疫重点开放实验室
关键词：
猪嗜血支原体;MSG1蛋白;酶标单抗;阻断ELISA
摘  要：
为确定猪嗜血支原体(M.suis)在临床的感染情况和进行流行病学调查提供检测方法,本研究以纯化的猪嗜血支原体重组MSG1蛋白和辣根过氧化酶标记的MSG1蛋白单抗建立了检测猪嗜血支原体的阻断ELISA方法。经筛选确定,抗原最佳包被浓度为0.5μg.mL-1,待检血清最佳稀释度为1∶1,作用时间为37℃1.5h,酶标单抗最佳稀释度为1∶10 000,作用时间为37℃1h。通过对100份M.suis阴性血清阻断ELISA检测结果进行统计学分析,确定本方法的判定标准为当阻断率PI≥23.71%时判为阳性,PI≤36.35%时判为阴性,23.71%
译  名：
Development of Blocking ELISA for Mycoplasma suis with Peroxidase-Labelled Monoclonal Antibody Against MSG1 Protein
作  者：
ZHANG Chang-ying,LI Yu-feng,JIANG Ping (Key Laboratory of Animal Diseases Diagnostic and Immunology of Ministry of Agriculture, College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)
关键词：
Mycoplasma suis;MSG1 protein;monoclonal antibody;blocking ELISA
摘  要：
To provide a serological method for detecting antibodies against Mycoplasma suis(M.suis),a blocking ELISA was developed based on a monoclonal antibody(MAb) to M.suis-MSG1 protein.The conditions for each step were optimized.The optimal concentration of the coating antigen was 0.5 μg·mL-1;sera samples diluted 1 fold and incubated 1.5 h at 37 ℃;MAb-HRP dilution is 10 000 fold and incubated 1 h at 37 ℃.The blocking results of one hundred M.suis negative sera samples were statistically analyzed,and the cutoff of blocking ELISA was determined that the samples presenting a percentage inhibition of ≥ 23.71% were considered positive;samples with a calculated percentage inhibition of ≤ 36.35% were rated negative and those presenting a blocking effect between 23.71% and 36.35% were considered inconclusive.The Blocking ELISA proved to be specific,sensitive and it showed high reproducibility and low variability.This method will be useful in clinical detection and epidemiological study on M.suis.

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