建立了一种快速检测单增李斯特菌的聚合酶链式反应-核酸层析(PCR-NALF)方法,该方法通过一对标记生物素和地高辛的引物,对提取自样品中的DNA进行扩增,然后用包被亲和素和抗地高辛抗体的层析试纸条对扩增产物进行检测,并通过肉眼可见的红色条带进行结果判读,从而替代电泳实现便捷检测。在特异性试验中,其他李斯特菌和常见致病菌均呈现阴性反应,与电泳结果一致。与传统微生物检测法(GB/T 4789.30-2008)相比无显著性差异(χ~2=0,P>0.05)。

This article presents a new Polymerase Chain Reaction-Nucleic Acid Lateral Flow(PCR-NALF) method for the rapid detection of Listeria monocytogenes(LM) in food.Following isolation of DNA from the samples, PCR with two specific primers,one labeled with biotin and the other with digoxigenin,respectively,was performed. After applying the amplified product into a lateral flow strip that is coated with avidin and anti-digoxigenin antibodies, the test result can be interpreted according to the visible lines on the strip.The specifics were studied by testing a range of Listeria strains and other food relevant microorganisms.PCR products from other nonpathogenic Listeria,other microorganisms and the primer control(PCR without template DNA) were all negative,so this method can exactly differentiate LM from other common bacteria.And there is no observed difference(x~2 =0,P >0.05) when compared with classical microbiological detection method(GB/T 4789.30-2008).As a rapid,convenient,sensitive and specific assay method,the PCR-NALF can provide an effective means for LM detection in food samples.