Prokaryotic Expression of PEB1A Protein of Campylobacter jejuni and Establishment of an Indirect ELISA

In order to develop an indirect ELISA method to detect Campylobacter jejuni antibody, PEB1A gene of Campylobacter jejuni was cloned and amplified by PCR, the prokaryotic expression vector pET32a-PEBIA was constructed, and then transferred into the expression strain E.coli BL21 (DE3), and obtained about 47 ku of soluble protein. Western blotting result showed that the expressed recombinant protein had good biological activity, an indirect ELISA method for detecting antibody against Campylobacter jejuni was developed using expressed PEB1A protein as coating antigen,and detected its specificity, sensitivity, repeatability, respectively. The results showed that the established method for detection of Campylobacter jejuni antibody critical value was 0.3424. This method only specifically reacts with Campylobacter jejuni positive sera, and had no cross-reactivity with other antiserum and strong specificity. In addition, the coefficients of variations in both inter-and intra-assay were less than 5% indicating that it had good repeatability and stability. The establishment of this method could be applied to the rapid detection of Campylobacter jejuni in serum, and provided basis for further prevention and control of Campylobacter jejuni diarrhea.