Prokaryotic Expression of Anaplasma marginale MSP2 Protein and Development of Indirect ELISA for Antibody Detection
In order to establish a serological assay for Anaplasma marginale,this study was conducted based on the gene sequence of A.marginale MSP2 collected by GenBank(Accession number:EU526889),the recombinant plasmid pET28a-MSP2 was constructed,and the pET28a-MSP2 protein was obtained and purified by prokaryotic expression.The purified protein was used as antigen to establish an indirect ELISA method.The results showed that the indirect ELISA method established in this study showed that the optimal concentration of antigen coated was 8μg/mL,the optimal serum dilution was 1:400,and the optimal working concentration of the secondary antibody was 1:2 000,which showed good specificity,sensitivity and repeatability.In this study,the MSP2 protein of A.marginale was successfully expressed and purified,and an indirect ELISA method for A.marginale antibody was established,which provided certain reference value for the monitoring,diagnosis for A.marginale.
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