大肠杆菌O抗原定型血清标定用抗原的制备

The Preparation of E. coli’s Antigen which Calibrate the Titer of the Mono-Factor Stereotype Serum

关键词:  大肠杆菌  菌体抗原  单因子血清

Key words:  Escherichia coli  bacterial antigen  mono -factor stereotype serum

摘要:

为研制用于标定大肠杆菌菌体抗原(O抗原)单因子定型血清效价的抗原,以大肠杆菌参考菌株CVCC1345(O2)、CVCC1350(O7)、CVCC1414(O74)为菌种,经条件优化确定了抗原制备的工艺。检测结果表明,当菌液调整至OD_600nm值为0.147±0.026时,各型抗原的浓度为1×10₉CFU/mL。将其热处理后制备成反应抗原。按此吸光值各自制备3批抗原,3批抗原与血清的凝集价均分别一致,分别达到2₁₁(O2)、2₁₂(O7)及2₉(O74)。用180种大肠杆菌O抗原单因子定型血清对制备的抗原进行检测,制备的抗原仅与其对应型的血清发生特异性反应,与其余179种血清均不发生反应。研究结果显示,制备的抗原具有较好的稳定性及特异性,可以用于大肠杆菌菌体单因子定型血清效价的标定。

Abstract:

In order to develop antigen that can be used to calibrate the titer of E. coli bacteria’s single factor stereotype serum. In this study the E. coli reference strain CVCC1345 (O2), CVCC1350 (O7), CVCC1414 (O74) as a species, is determined by the conditions to optimize their production processes. Our test results show that the bacteria which will be prepared as the antigen after the heat treatment adjusted to OD600nm value of 0.147±0.026, the concentration of 1× 10₉CFU/mL. According to this absorbance value of each antigen, we have 3 batches of the prepared antigen. Three batches of antigen and serum agglutination titer are consistent respectively which reached 2₁₁ (O2), 2₁₂ (O7) and 2₉ (O74). We test the prepared antigen with 180 kinds of mono-factor stereotype serum of E. coli, only the prepared antigen of the corresponding type of serum could react specifically, the rest of the 179 kinds of serum not react. The results showed that the prepared antigen having better stability and specificity of E. coli bacteria can be used to calibrate the titer of the mono-factor stereotype serum.