大肠杆菌菌体O抗原单因子定型血清的制备

Preparation of Mono-specific Anti-O-antigen Serum of E.coli

关键词:  大肠杆菌  菌体抗原  单因子血清

Key words:  Escherichia coli  somatic antigen  mono-specific serum

摘要:

为研制大肠杆菌菌体O抗原单因子定型血清，以大肠杆菌参考菌株CVCC1345（O2）、CVCC1350（O7）、CVCC1414（O74）为菌种制备抗原免疫家兔，获得了定型原血清，并通过比较不同免疫程序及剂量优化了定型原血清的制备工艺。进而按照优化后的定型原血清制备工艺，以大肠杆菌参考菌株CVCC1343（O1）、CVCC1345（O2）、CVCC1350（O7）、CVCC1405（O64）、CVCC1414（O74）、CVCC1439（O100）、CVCC1442（O103）、CVCC1455（O116）、CVCC1466（O128）、CVCC3801（O154）为菌种制备抗原免疫家兔，获得了10种O抗原型定型原血清。将定型原血清分别与180种微量凝集试验抗原逐一反应，明确各原血清与非特异性抗原的交叉凝集价，通过用吸收抗原对原血清进行吸收及稀释的方法消除非特异性凝集，最终制备成大肠杆菌菌体O抗原单因子定型血清。血清质量评价结果表明，新制备的单因子定型血清性状为无色、淡黄色或黄色澄明液体，个别有少量絮状物；无菌生长；特异性良好；凝集效价为1︰2。研究结果显示，制备的大肠杆菌O抗原单因子定型血清具有较好的特异性，可以用于对临床分离大肠杆菌进行O抗原血清型鉴定。

Abstract:

Mono-specific anti-O-antigen serum of Escherichia coli (E.coli) was developed and prepared in this research. Firstly, antigens were prepared using E.coli reference strains of CVCC1345 (O2), CVCC1350 (O7) and CVCC1414 (O74) respectively and raw sera were obtained from rabbits immunized with the antigens above separately. Productive technology of raw serum was optimized by comparing different immune procedures and doses of the three antigens. Then ten raw sera were harvested using the optimized productive technology by immunizing rabbits with antigens prepared from 10 E.coli reference strains CVCC1343（O1），CVCC1345（O2），CVCC1350（O7），CVCC1405（O64），CVCC1414（O74），CVCC1439（O100），CVCC1442（O103），CVCC1455（O116），CVCC1466（O128），CVCC3801（O154）. Cross-agglutination was detected by reacting sera with 180 micro-agglutination test antigens respectively and eliminated by absorption with non-specific antigens and dilution. Finally mono-specific anti-O-antigen sera of E.coli were obtained. Quality evaluation of sera showed that newly prepared mono-specific anti-O-antigen sera were colorless, pale yellow or yellow clear sterilized liquid with good specificity. Only a few had a little floccule. Agglutination titer was 1∶2. All the results suggested that newly prepared mono-specific anti-O-antigen sera could be applied in serotype identification of clinical isolated E. coli strains with their good specificity.