【摘要】 目的制备抗霍乱弧菌O1群特异性单克隆抗体(McAb),并进行鉴定。方法以福尔马林灭活的O1群霍乱弧菌免疫BALB/c小鼠,应用细胞融合技术建立分泌抗O1群霍乱弧菌McAb的杂交瘤细胞株。对配对较好的6个细胞株分泌的McAb进行腹水效价、免疫球蛋白类及亚类、抗体亲和常数测定及凝集试验和稳定性试验。与细菌培养法对比检测208份临床标本。结果6个细胞株分泌的McAb腹水效价均为105,均为IgM;抗体亲和常数为1.84×105～5.38×107;且均与O1群霍乱弧菌菌株发生凝集反应,与霍乱弧菌O139群、大肠杆菌、出血性大肠杆菌O157∶H7、副溶血弧菌、麦弧菌、河弧菌、结肠炎耶尔森菌、普通变形杆菌、痢疾杆菌、伤寒杆菌及各型副伤寒杆菌均不发生凝集反应。6个分泌McAb的细胞株连续培养15周,经液氮冻存12个月后复苏,仍能稳定分泌抗体;与细菌培养法对比检测208份临床标本,特异性和灵敏度均达100%。结论制备的McAb具有较高的特异性,有可能用于开发霍乱弧菌O1群的检测试剂。更多还原

【Abstract】 Objective To prepare and identify the monoclonal antibody (McAb) against Vibrio cholerae group O1. Methods The hybridoma cell strains secreting McAbs were established by fusing SP2/0 cells with the spleen cells of BALB/c mice immunized with V. cholerae group O1 inactivated by formalin. Six cell strains were screened by colloidal gold matching test, and the secreted McAbs were tested for titer in ascites, class and subclass of IgG and antibody affinity constant, then subjected to agglutination and stability tests. A total of 208 clinical specimens were determined with the prepared McAbs, and the results were compared with those by bacterial culture. Results All the McAbs secreted by 6 hybridoma cell strains were IgM, each at a titer of 105 in ascites, and their antibody affinity constants ranged from 1. 84 × 105 to 5. 38 × 107. All the McAbs showed agglutination reaction with V. cholerae group O1, but no reaction with V. cholerae group O139, E. coli, enterohemahagic E. coli O157 ∶ H7, V. parahaemolyticus, V. metchnikovii, V. fluvialis, Y. enterocolitica, P. vulgaris, Sh. dysenteriae, Sal. typhi or Sal. paratyphi A, B or C. After the 6 hybridoma cell strains were cultured continuously for 15 weeks and stored in frozen for 12 months, McAbs were still secreted stably. Both the specificity and sensitivity of the prepared McAbs in determination of 208 clinical specimens were 100%. Conclusion The McAbs secreted by established 6 hybridoma cell strains showed good specificity and might be used for the development of diagnostic kit for V. cholerae group O1 infection.更多还原