利用 MSG1 蛋白建立一种新型阻断 ELISA 检测方法用于猪肺炎支原体感染的诊断。
Use of MSG1 protein in a novel blocking ELISA for the detection of Mycoplasma suis infection.
机构信息
Key Laboratory of Animal Diseases, Diagnostics and Immunology, Ministry of Agriculture, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
出版信息
Vet J. 2012 Aug;193(2):535-8. doi: 10.1016/j.tvjl.2011.12.017. Epub 2012 Jan 27.
A blocking ELISA based on the monoclonal antibody (MAb) 1A7 was developed to detect antibodies against Mycoplasma suis. The MAb was produced by immunising BALB/c mice with recombinant MSG1 protein (rMSG1) from M. suis expressed in E. coli. Following identification by Western blotting, the MAb was purified and labelled with horseradish peroxidase. The parameters of the ELISA were optimised, and the cut-off value determined as 36.35% by analysing the percentage inhibition of M. suis negative serum. The sensitivity and specificity of the ELISA were 92% and 100%, respectively. In repeatability tests, the intra- and inter-batch variation coefficients were <10%. The results suggest this blocking ELISA is specific, sensitive and reproducible, and will be a valuable tool in the serodiagnosis of M. suis infection in pigs.
建立了一种基于单克隆抗体（MAb）1A7 的阻断 ELISA 来检测抗猪支原体的抗体。该 MAb 通过用大肠杆菌中表达的重组 MSG1 蛋白（rMSG1）免疫 BALB/c 小鼠产生。通过 Western blot 鉴定后，将 MAb 纯化并用辣根过氧化物酶标记。通过分析猪支原体阴性血清的抑制率，优化了 ELISA 的参数，并将截断值确定为 36.35%。该 ELISA 的灵敏度和特异性分别为 92%和 100%。在重复性试验中，批内和批间变异系数均<10%。结果表明，这种阻断 ELISA 具有特异性、敏感性和重现性，将成为猪支原体感染血清学诊断的有价值工具。
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