摘要
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 a quick checking method of enterotoxins genes in food-borne Listeria monocytogenes by multiplex polymerase chain reaction （MPCR） and denaturing high-performance liquid chromatography （DHPLC）. Methods Primers for detection of hly, prfA, inlA and inlB were designed and PCR system was optimized. Products were detected by DHPLC for quick detection. Thirty-two strains were tested with PCR method, sensitivity was determined with various concentration of standard strains. Results The peak order of PCR products was inlB, hly, inlA, prfA, and amplified fragment size was 146, 210, 255 and 388 bp respectively. This method has good specificity, and the lowest amount of detecting was 280 cfu/ml. Conclusion This method can well meet the requirements of actual food microbe testing.
出处
《中国食品卫生杂志》
北大核心
2014年第2期141-145,共5页
Chinese Journal of Food Hygiene
