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目的建立单增李斯特菌的多个靶基因快速检测手段,提高检测的准确性。方法根据单增李斯特菌4个毒力基因(hly、prfA、inlA、inlB)设计引物,通过优化引物浓度和引物组合,进行多重PCR扩增,产物经变性高效液相色谱(DHPLC)进行快速检测。结果出峰顺序依次为inlB、hly、inlA、prfA,扩增片段大小为146、210、255、388 bp,此方法具有良好的特异性,灵敏度可达到280 cfu/ml。结论本方法可以满足实际工作中食品微生物检测的要求。
Objective To establish rapid detection of multiple target genes of Listeria monocytogenes and improve the accuracy of detection. Methods Primers were designed according to four virulence genes of Listeria monocytogenes (hly, prfA, inlA, inlB). Multiplex PCR amplification was performed by optimizing the primer concentration and primer combination. The product was rapidly denatured by high performance liquid chromatography (DHPLC) Detection. The results showed that the peak order of inlB, hly, inlA and prfA was 146,210,255,388 bp. This method has good specificity and sensitivity of 280 cfu / ml. Conclusion This method can meet the requirements of food microbiological testing in practical work.