[使用聚合酶链反应（PCR）方法鉴定牛奶中的单核细胞增生李斯特菌]

[Use of PCR methods for identification of Listeria monocytogenes in milk].

作者信息

Wieckowska M, Kotłowski R, Kur J, Rudnicka W

机构信息

Zakład Biologii Infekcyjnej Instytutu Mikrobiologii i Immunologii, Uniwersytetu Lódzkiego.

出版信息

Med Dosw Mikrobiol. 1998;50(3-4):251-7.

PMID:10222740

Abstract

The aim of this work was to estimate the limit of Listeria monocytogenes cfu in polymerase chain reaction (PCR) for a DNA fragment of listeriolysine O (hly A) gene. The PCR method, with used primers selected in areas of the listeriolysin O gene, allows to differentiate L. monocytogenes strains from other Listeria species. The amplified fragment (456 bp) of hly A gene was obtained for all strains L. monocytogenes and no other Listeria species. The PCR method with the selected primers allowed to detect 50-500 cfu L. monocytogenes/ml suspended in water or milk. Among 20 samples of raw milk from cows, 10 samples contained > 50 cfu L. monocytogenes/ml. Obtained results indicate that the PCR assay of L. monocytogenes identification is technically simple and may be conduct with minimal time. So, it could be recommended as quick diagnostic method in identification L. monocytogenes in milk.

摘要

这项工作的目的是估计聚合酶链反应（PCR）中用于李斯特菌溶血素O（hly A）基因DNA片段的单核细胞增生李斯特菌菌落形成单位（cfu）的限度。采用在李斯特菌溶血素O基因区域选择的引物的PCR方法，能够区分单核细胞增生李斯特菌菌株与其他李斯特菌属物种。所有单核细胞增生李斯特菌菌株均获得了hly A基因的扩增片段（456 bp），而其他李斯特菌属物种均未获得。采用所选引物的PCR方法能够检测出悬浮于水或牛奶中的50 - 500 cfu/ml的单核细胞增生李斯特菌。在20份奶牛生鲜乳样品中，有10份样品的单核细胞增生李斯特菌含量>50 cfu/ml。所得结果表明，单核细胞增生李斯特菌鉴定的PCR检测技术简单，且可在最短时间内完成。因此，它可作为牛奶中单核细胞增生李斯特菌鉴定的快速诊断方法加以推荐。