小肠结肠炎耶尔森菌O∶9血清型特异性单克隆抗体筛选及其应用

Screen and Its Amplification of Specific mAb of Yersinia enterocolitica Serotype O∶9

目的利用杂交瘤技术筛选出能分泌小肠结肠炎耶尔森菌O∶9血清型特异性单克隆抗体的细胞株,用于该型菌株的鉴定。方法采用ELISA方法筛选克隆株,并用玻片凝集法进行单克隆抗体的特异性鉴定。结果筛选到3株能分泌小肠结肠炎耶尔森菌O∶9血清型特异性单克隆抗体的杂交瘤细胞株,分泌的抗体与经鉴定的18株小肠结肠炎耶尔森菌O∶9血清型菌株呈阳性反应,而与其他血清型小肠结肠炎耶尔森菌及常见肠道致病菌无交叉凝集反应;应用于小肠结肠炎耶尔森菌野生株分型鉴定时,能准确鉴定出其中的24株O∶9血清型菌株,与用日本进口分型血清鉴定、PCR毒力基因检测结果相同。结论筛选获得的细胞株产生的单克隆抗体,用于小肠结肠炎耶尔森菌O∶9血清型鉴定,具有很好的型特异性,避免了过去免疫血清制备中繁琐的抗原交叉吸收试验。

Objective To prepare specific mAb of Yersinia enterocolitica serotype O：9 and to detect its specificity. Methods Clones were screening by ELISA and the specificity was identified by slide agglutination. Results The mAbs of three cell strains reacted positively with all of the strains of Yersinia enterocolitica serotype O：9 and negatively with other serotypes of Yersinia enterocolitica and other enteric-pathogenic bacterium. The results were accordance with the result tested by Japanese typing serum. Conclusion The three cell strains of mAb of Yersinia enterocolitica serotype O：9 got from this study can be used to identify strains of Yersinia enterocolitica serotype O： 9. These mAbs shew good serotype specificity. The cross-agglutinating and adsorption assay are unnecessary comparing with the preparation of rabbit immune serum.