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单核细胞增生李斯特氏菌PCR-DHPLC检测新技术的建立

作  者:
曹际娟;闫平平;徐君怡;郑秋月;马惠蕊;谢明杰
单  位:
辽宁师范大学;辽宁出入境检验检疫局
关键词:
DHPLC;单核细胞增生李斯特氏菌;fimY基因
摘  要:
应用PCR结合变性高效液相色谱(DHPLC)技术建立食品中单核细胞增生李斯特氏菌fimY的快速检测方法。根据单核细胞增生李斯特氏菌prfA和hlyA基因序列的特点设计特异性引物,PCR扩增的产物经DHPLC技术进行快速检测。以单核细胞增生李斯特氏菌等61株参考菌株做特异性试验;单核细胞增生李斯特氏菌菌株稀释成不同梯度,进行灵敏度试验。试验结果表明该方法具有很好的特异性,灵敏度较高,检测低限可达到为181CFU/ml。可以快速、准确检测单核细胞增生李斯特氏菌,是食品中致病菌快速检测的新技术。
译  名:
Study of Identification of Listeria monocytogenes with PCR-DHPLC
作  者:
Cao Jijuan~1 Yah Pingping~2 Xu Junyi~1 Zheng Qiuyue~(1,3)Ma Huirui~1 Xie Mingjie~2 (1 Liaoning Entry-Exit Inspection & Quarantine Bureau of P.R.China,Dalian 116001; 2 Liaoning Normal University,Dalian 116021;3 Shenyang Agricultural University,Shenyang 110161)
关键词:
DHPLC;;Listeria monocytogenes;;prfA and hlyA gene
摘  要:
To identify the prfA and hlyA gene in Listeria rnonocytogenes,a PCR-DHPLC assay was performed in this study.Primers specific for the prfA and hlyA gene in Listeria monocytogenes of food were selected to conduct the DHPLC assays.The specific testing was performed with Listeria monocytogenes and 61 strains,and various grads of Listeria monocytogenes was used for the sensitivity testing.The good specificity was found in the study.Also,the method showed nicer sensitivity with the lowest amount of detecting was 181 CFU/mL,so it can detect and identify Listeria monocytogenes quickly and correctly.The DHPLC could be a new method for detecting food pathogens.

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