摘要
研究获得纯化抗大肠杆菌O157:H7IgY抗体,经检测10mg/ml纯化IgY抗体的效价为1:320;以大肠杆菌O157:H7免疫新西兰大耳白兔,获得兔抗大肠杆菌O157:H7IgG抗体,效价达1:25600。以兔抗大肠杆菌O157:H7IgG抗体稀释3200倍作为捕获抗体,抗大肠杆菌O157:H7IgY抗体为检测抗体建立双抗夹心ELISA方法检测大肠杆菌O157:H7,正交试验分析表明,捕获抗体于37℃包被2h、不封闭、抗原与捕获抗体于37℃结合2h、检测抗体浓度为0.25mg/ml、与抗原于37℃结合1h为最优反应条件。该方法对纯培养菌液检出限为105CFU/ml,具有良好的敏感性及特异性。染菌样品经在EC增菌液中选择性培养后进行双抗夹心ELISA检测,接种量为0.1～1CFU/g(ml)的样品在培养12h后可检出阳性反应,1～10CFU/g(ml)的样品在培养8h后可检出阳性反应。
In this paper, anfi-E.coli O157：H7 IgY was purified, and the titer of 10mg/ml purified IgY was 1：320. The anti- E.coli O 157：H7 IgG was available by injection New Zealand rabbit with E.coli O 157 ：H7 antigen, and the liter was 1：25600.Together with the IgY and IgG specific to E. coli O157, a double-antibody sandwich-ELISA was developed with IgG diluted 3200 fold as the capture antibody, and IgY as the detection antibody. Besides, the results of cross-experimentation of sandwich-ELISA described： ① IgG coating in 37℃ for 2h, ② reaction of IgG with antigen in 37℃ for 2h, and ③ concentration of IgY as 0.25mg/ml, and reaction of IgY with antigen in 37℃ for lb. The detection limit of this method is 10^5CFU/ml, and has no cross reactions with other bacteria. With enrichment procedure in 41℃ and 200 r/min, E. coli O 157：H7 recovers well from inoculated food samples. The results showed that the samples are detected after being cultured 12h at 0.1 - 1CFU/g（ml） and 8h at 1 10CFU/g（ml） respectively.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2007年第1期171-175,共5页
Food Science
关键词
大肠杆菌O157:H7:双抗夹心ELISA
检测
E. coli O 157： double-antibody sandwich ELISA（enzyme linked immunosorbent assay）： detect
