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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 screen the cell strain secreting Yersinia enterocolitica O: 9 serotype specific monoclonal antibody by hybridoma technique and to identify the strain. Methods Clonal strains were screened by ELISA and the specificity of monoclonal antibodies was determined by slide agglutination. Results Three hybridoma cell lines secreting Yersinia enterocolitica O: 9 serotype-specific monoclonal antibodies were screened. The secreted antibodies were separated from the identified Yersinia enterocolitica O: 9 serum Type strains positive reaction, and with other serotype Yersinia enterocolitica and common intestinal pathogens no cross-agglutination reaction; used in the identification of Yersinia enterocolitica wild type identification can accurately identify Among them, 24 isolates of O: 9 serotypes were identified by PCR-based virulence analysis with the imported type sera from Japan. Conclusion Monoclonal antibodies produced by the selected cell lines were used in the identification of Yersinia enterocolitica O: 9 serotypes, which showed good specificity and avoided the cumbersome antigen cross-absorption test in the past.