Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (21): 4508-4515.doi: 10.3864/j.issn.0578-1752.2011.21.020

• VETERINARY SCIENCE • Previous Articles     Next Articles

Establishment of ELISA Based on Recombinant Protein MSG1 for the Detection of Mycoplasma suis Antibody

 LIU  Yong-Xia, LIU  Jian-Zhu, CHENG  Zi-Qiang, ZHOU  Dong, WANG  Zhen-Yong, LIU  Hai-Tao, WANG  Lin, CHAI  Tong-Jie   

  1. 1.山东农业大学动物医学院/山东省动物生物技术与疫病防治重点实验室，山东泰安 271018
  • Received:2011-03-04 Online:2011-11-01 Published:2011-09-16

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Abstract: 【Objective】 An enzyme-linked immunosorbent assay (ELISA) system based on the polyclonal antibody against the adhesion protein MSG1 which was coded by the g1 gene of Mycoplasma suis(M. suis) was established, and was assessed as an effective method for detection of M. suis in swine. 【Method】 The MSG1-pET28c_E.coli BL21 was induced to express the MSG1protein of M. suis, and broken by ultrasound, the aim protein was purified by nickel affinity chromatography. The immunogen that was made of this purified recombinant protein was vaccinated to the mice to produce the polyclonal antibody. The polyclonal antibody and the recombinant protein were used to build an ELISA to detect M. suis, and the sensitivity, specificity, coincidence and repeatability of this method was analyzed, respectively. 【Result】 The polyclonal antibody was obtained whose titer had reached an adequate level. The infection rate of M. suis was 46.25% through the built ELISA method, the positive samples of them could hinder the polyclonal antibody from integrating with the MSG1protein markedly and didn’t react with some other antigens. The sensitivity and specificity values reached 95.92% and 97.06%, respectively, and a higher detection rate was confirmed through the ELISA method comparing with the IHA method. 【Conclusion】 The results show that the antigenicity of the protein MSG1 was satisfactory. This ELISA system had a higher sensitivity, specificity, coincidence and stability for the serodiagnosis of M. suis. A desirable result would be obtained from the detection of M. suis.

Key words: Mycoplasma suis, MSG1, ELISA, establishment

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