摘要
建立大肠杆菌O157：H7荧光纳米颗粒免疫层析法。方法：首先将荧光纳米颗粒与大肠杆菌O157：H7单抗共价偶联，制成大肠杆菌O157：H7单抗-荧光纳米颗粒偶联物。用该偶联物代替金标颗粒，以双抗体夹心作为反应模式来制备大肠杆菌O157：H7免疫层析试纸条。在紫外灯下观察免疫层析试纸条上经免疫反应而产生的质控线和检测线上的荧光信号，并利用荧光强度来半定量检测大肠杆菌O157：H7。结果：用所制备的试纸条对11属24种54株菌进行检测，所有27株大肠杆菌O157：H7的检测结果呈阳性．其它非大肠杆菌O157菌株检测结果呈阴性。用该试纸条检测人工污染的鸡肉样品。灵敏度为2．7×10^4CFU／mL。通过观察检测线的有无。可对大肠杆菌O157：H7进行半定量检测。分别用所制备的大肠杆菌O157：H7免疫层析试纸条和标准法检测57份样品，2种方法的总符合率为80．7％。结论：大肠杆菌O157：H7荧光纳米颗粒检测试纸条的研制成功，为大肠杆菌O157：H7的快速检测提供了一个极好的检测方法．便于野外检测的开展．具有广阔的应用前景。
Establish a rapid method for the detection of E. eoli O157：H7. Method：Fluorescent nanopartieles were covalently conjugated with monoclonal antibodies against E. eoli O157 ：H7 and were employed in the development of lateral flow strip assay. The sensitivity, specificity and semi - quantitative of the lateral flow strip were evaluated. Results The sensitivity that detected spiked chicken sample was 2.7 104CFU/mL. The LAMP assay correctly identified 27 E. eoli O157：H7 strains, but did not detect 14 non - E. coli O157 ：H7 strains. With this method, semi -quantitative detection of E. coli O157 ：H7 can be finished in less than 15rain. This method was compared with standard method （SN/T 0973 -2000） and the agreement rates are 80.7%. Conclusion ：This lateral flow strip assay is sensitive, simple, rapid method without the need of special apparatus, so it is of good practical value for the detection of SN/ T 0973 - 2000.
基金
国家质检总局科技计划项目（2006SZK14）
