摘要
目的建立双重荧光定量PCR方法,快速检测沙门菌和单核细胞增生李斯特菌。方法通过设计特异性引物和探针,扩增沙门菌的fimY基因和单核细胞增生李斯特菌的hly基因,采用倍比梯度稀释法检测该体系的灵敏度,以另外7株肠道致病菌评价检测体系的特异性;建立了沙门菌和单核细胞增生李斯特菌感染小鼠的检测模型以验证方法的适用性。结果建立了同时检测沙门菌和单核细胞增生李斯特菌的双重荧光定量PCR方法,从DNA提取到检测完毕仅需2.5 h。检测两种病原菌的灵敏度分别为11和12.8 copies/μl,特异性为100%,符合率为93.3%。结论该法缩短了检测时间,并有良好的灵敏性和特异性,在疾病防控及食品卫生行业中很有应用前景。
Objective To develop a duplex real-time PCR array for rapid detection of Salmonella and Listeria monocytogenes.Methods The specific primers and probes were designed to amplify the fimY gene of Salmonella and the hly gene of Listeria monocytogenes.The sensitivity of the system was detected by a multiple proportional dilution method.In order to examine the specificity of the system,other seven intestinal bacteria strains were assayed simultaneously.Salmonella and Listeria monocytogenes infected mice model were reproduced for evaluating the applicability of the method.Results A highly sensitive and specific duplex real-time PCR assay for the detection of Salmonella and Listeria monocytogenes was established.The sensitivity was 11 copies/μl for Salmonella and 12.8 copies/μl for Listeria monocytogenes.The specificity was 100% and the accuracy was 93.3%.The whole detection procedure can be finished within 2.5 h.Conclusion The duplex real-time PCR detection method is efficient in detecting Salmonella and Listeria monocytogenes in foods with good sensitivity and specificity.Due to the simultaneous detection of these two pathogens,the detection time was reduced significantly.There is a good prospect of this method applying in disease prevention and food industry.
出处
《中国食品卫生杂志》
北大核心
2011年第4期289-292,共4页
Chinese Journal of Food Hygiene
基金
质检公益性行业科研专项(2007GYJ0252007GYJ023)
