[用于快速检测模拟牛奶样本中单核细胞增生李斯特菌的实时聚合酶链反应检测法]
[Real-time PCR assay for rapid detection of Listeria monocytogenes in simulated milk specimens].
作者信息
Wang Ping, Yuan Fei, Yang Hairong, Zhao Yongsheng, Hu Yue, Zhao Guiming, Chen Ying
机构信息
Chinese Academy of Inspection and Quarantine, Beijing 100123, China.
出版信息
Wei Sheng Yan Jiu. 2011 Nov;40(6):765-8.
OBJECTIVE
To establish a real-time PCR assay for the rapid detection of Listeria monocytogenes in simulated milk specimens.
METHODS
Based on part fragments of hlyO gene, a pair of primers and Taq-Man probe were designed for quantitative detection of L. monocytogenes. The specificity of the primers and probe were tested by using different L. monocytogenes strains and other common pathogenic bacteria.
RESULTS
L. monocytogenes strains were positive in the detection and other tested strains were negative. The sensitivity of assay was 9 copies per PCR reaction.
CONCLUSION
The specificity and sensitivity of Taq Man real-time PCR technology for detecting L. monocytogenes in simulated dairy specimens were high, and the assay could be completed within 1.5 h. This method could be used to detect other food samples contaminated by L. monocytogenes and identify the cause of food-borne Listeriosis outbreaks.
目的
建立一种实时荧光定量聚合酶链反应（PCR）检测方法，用于快速检测模拟牛奶样本中的单核细胞增生李斯特菌。
方法
基于hlyO基因的部分片段，设计一对引物和Taq-Man探针用于单核细胞增生李斯特菌的定量检测。通过使用不同的单核细胞增生李斯特菌菌株和其他常见病原菌检测引物和探针的特异性。
结果
单核细胞增生李斯特菌菌株检测呈阳性，其他检测菌株呈阴性。该检测方法的灵敏度为每个PCR反应9个拷贝。
结论
Taq Man实时荧光定量PCR技术检测模拟乳制品样本中单核细胞增生李斯特菌的特异性和灵敏度高，检测可在1.5小时内完成。该方法可用于检测其他被单核细胞增生李斯特菌污染的食品样本，并确定食源性李斯特菌病暴发的原因。
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