采用免疫磁珠分离结合聚合酶链反应快速检测火腿样品中的单核细胞增生李斯特菌。

Rapid detection of Listeria monocytogenes in ham samples using immunomagnetic separation followed by polymerase chain reaction.

作者信息

Hudson J A, Lake R J, Savill M G, Scholes P, McCormick R E

机构信息

Institute of Environmental Science and Research (ESR) Ltd, Christchurch Science Centre, PO Box 29-181, Ilam, Christchurch, New Zealand.

出版信息

J Appl Microbiol. 2001 Apr;90(4):614-21. doi: 10.1046/j.1365-2672.2001.01287.x.

DOI:10.1046/j.1365-2672.2001.01287.x

PMID:11309074

Abstract

AIMS

To develop a 24-h system for the detection of Listeria monocytogenes in ham.

METHODS AND RESULTS

An immunomagnetic separation (IMS) of bacteria directly from ham followed by extraction of DNA and detection using a new multiplex polymerase chain reaction (PCR) was used. The PCR method used one primer pair targeted at the listeriolysin O gene of L. monocytogenes and the other pair for a region of the 23S rRNA genes of Listeria, giving products of 706 and 239 bp, respectively. The combined IMS/PCR was calculated to be capable of detecting as few as 1.1 L. monocytogenes cells g-1 in a 25-g ham sample.

CONCLUSION

The process produced acceptable results, but the IMS step is the main barrier to further improvement of sensitivity. The DNA isolation was the most time-consuming step in the process.

SIGNIFICANCE AND IMPACT OF THE STUDY

A 24-h test for the presence of L. monocytogenes will be useful to the food industry and significantly assist in the timely investigation of outbreaks.

摘要

目的

开发一种用于检测火腿中单核细胞增生李斯特菌的24小时检测系统。

方法与结果

采用直接从火腿中进行细菌免疫磁珠分离（IMS），随后提取DNA并使用新型多重聚合酶链反应（PCR）进行检测。该PCR方法使用一对引物靶向单核细胞增生李斯特菌的溶血素O基因，另一对引物针对李斯特菌23S rRNA基因的一个区域，分别产生706和239 bp的产物。经计算，联合IMS/PCR能够在25 g火腿样品中检测到低至1.1个单核细胞增生李斯特菌细胞/g。