毛细管电泳法快速分离和检测肠毒性大肠杆菌

 建立了快速分离和检测引起仔猪腹泻的肠毒性大肠杆菌K88、K99和 987P细菌细胞的毛细管区带电泳方法 ,并进行了腹泻仔猪粪便中肠毒性大肠杆菌的应用检测分析。结果表明 ,在电泳缓冲液为 0 0 5mol/LNa2 CO3 NaHCO3(pH 9 9)、分离电压为 1 4 1kV、检测波长为 2 1 0nm的电泳条件下 ,E coliK88、K99和 987P的细胞分别具有单一、稳定的特征谱峰 ,其保留时间的相对标准偏差RSD≤ 0 9% ;在确定的实验条件下 ,实现了腹泻仔猪粪便中肠毒性大肠杆菌的快速检测分析 ,发现将出生 5d～ 6d的腹泻仔猪粪便引入培养基中增殖后主要检出K88。

Rapid Separation and Determination of Enterotoxigenic E.coli by Capillary Zone Electrophoresis

Rapid separation and determination of enterotoxigenic E. coli K88, K99 and 987P with intact cells using capillary zone electrophoresis (CZE) are described. The CZE was performed in a running buffer of 0.05 mol/L Na2CO3-NaHCO3 (pH 9.9), under applied voltage of 14.1 kV and detected by an on-line UV at 210 nm. Results show that K88, K99 and 987P were separated completely. Moreover, each strain presented a characteristic peak and a reproducible retention time (RSD < or = 0.9%). The successful analysis of enterotoxigenic E. coli in the excrement of diarrhea piglets is also discussed. K88 and K99 were detected in the excrement of diarrhea piglets born in 5 d - 6 d, and 30 d - 35 d respectively, and nothing was detected in the excrement of diarrhea piglets born in about 60 d.