胞内劳森菌LsaA蛋白的原核表达及间接ELISA抗体检测方法的建立

doi:10.11843/j.issn.0366-6964.2018.04.016

Abstract

Abstract:

In order to establish a method for detecting specific antibody against Lawsonia intracellularis, the codon-optimized LsaA gene encoding surface protein LsaA of L. intracellularis was cloned into pET32a (+) vector, and the expression of LsaA protein was induced in E. coli host BL21 (DE3). After optimization of a series of conditions, an indirect ELISA for detecting antibodies against L. intracellularis was established by using purified LsaA protein as coating antigen. The results showed that the anti- L. intracellularis antibodies were detected specifically by the established ELISA, and there were no cross-reactions with the antibodies against common diarrhea-causing pathogens such as E. coli, Salmonella choleraesuis, porcine epidemic diarrhea virus and transmissible gastroenteritis virus. Serum samples from fecal L. intracellularis-positive pigs which were identified by PCR, were also positive for anti- L. intracellularis antibodies detected by the established ELISA. The ELISA has good sensitivity and repeatability. Anti- L. intracellularis positive serum with 1:800 dilution was still ELISA positive. The intra-and inter-assay coefficient of variation was 0.352%-2.752% and 0.877%-3.000%, respectively. Seroepidemiological prevalence of L. intracellularis infection in partial pig farms in Henan province was investigated by using the established ELISA. The results indicated that all the tested pig farms were anti- L. intracellularis antibody positive with positive rate of 13.3% to 57.1%, with average positive rate of 30.6%. The results showed that the established ELISA in this study can be used for the detection of antibodies against L. intracellularis, and provides a method for the monitoring and epidemiological investigation of L. intracellularis.

CLC Number:

S852.61

WU Yan-yang, YANG Dong-dong, GAO Dong-sheng, LI Yong-tao, CHANG Hong-tao, WANG Chuan-qing, ZHAO Jun. Prokaryotic Expression of LsaA Protein of Lawsonia Intracellularis and Development of a LsaA-based Indirect ELISA for Antibody Detection[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(4): 786-793.

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Prokaryotic Expression of LsaA Protein of Lawsonia Intracellularis and Development of a LsaA-based Indirect ELISA for Antibody Detection

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