基于OMP18的B细胞抗原表位的空肠弯曲菌ELISA检测方法的建立

目的 以OMP18的B细胞抗原表位多肽为包被抗原,建立检测空肠弯曲菌感染的ELISA方法。方法 以不同浓度梯度(0.1,1,10 μg/mL)OMP18的B细胞抗原表位多肽进行包被,以空肠弯曲菌全菌兔抗IgG为一抗,HRP标记的羊抗兔抗体IgG为二抗,检测对原浓度1 mg/mL的不同稀释度(1:10,1:100,1:1 000)抗体IgG水平。分别以空肠弯曲菌感染兔血清、健康兔血清及沙门菌感染兔血清为一抗,1:3 000稀释的HRP标记的羊抗兔IgG为二抗,比较各抗原表位多肽的免疫特异性。结果 OMP18的B细胞抗原3个表位多肽在稀释度在1:1 000、1:4 000、1:16 000、1:64 000时免疫反应性均有明显增高。其中稀释度在1:1 000时增高最明显。OMP18的B细胞抗原表位多肽具有免疫特异性。结论 用OMP18的B细胞抗原表位多肽作为包被抗原对空肠弯曲菌感染的血清进行ELISA检测,免疫反应性高,具有免疫特异性。

Establishment of ELISA method for detecting Campylobacter jejuni based on the B-cell antigen epitope of OMP18

This study aims to establish an ELISA method for detection of Campylobacter jejuni infection by using the B-cell antigen epitope polypeptide of OMP18 as the coating antigen. Using the B-cell epitope peptides with different concentrations (0.1,1 and 10 μg/mL) as the coating antigens, the rabbit anti-Campylobacter jejuni IgG antibody as the primary antibody and the HRP-labeled goat anti-rabbit IgG antibody as the secondary antibody, the researchers detect the levels of IgG antibody after different dilutions (1:10, 1:10 and 1:1 000) based on its original concentration of 1 mg/mL. The researchers also compare different immunological specificities of epitope peptides via using the serum from a healthy rabbit, the serums respectively from the rabbits infected with Campylobacter jejuni and Salmonella as the primary antibody as well as the HRP-labeled goat anti-rabbit IgG antibody(1:3 000) as the secondary antibody. The immunoreactivity of three B cell antigen epitope polypeptides of OMP18 significantly increase in the dilutions of 1:1 000,1:4 000 and 1:16 000 and 1:64 000,while it finds the most significant increase in a 1:1 000 dilution.Therefore, the B-cell epitope peptides of OMP18 indeed features an immunological specificity. And the study indicates that the ELISA is a desirable method capable of detecting Campylobacter jejuni infection by using OMP18 B cell antigen epitope polypeptide as the coating antigen with a high immune response and a specific immunity.