通过聚合酶链反应（PCR）检测来自伊氏李斯特菌的一个假定的N - 乙酰胞壁酸酶基因有助于对其进行快速鉴定。

PCR detection of a putative N-acetylmuramidase gene from Listeria ivanovii facilitates its rapid identification.

作者信息

Liu Dongyou, Ainsworth A Jerald, Austin Frank W, Lawrence Mark L

机构信息

Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, P.O. Box 6100, Mississippi, MS 39762, USA.

出版信息

Vet Microbiol. 2004 Jun 21;101(2):83-9. doi: 10.1016/j.vetmic.2004.03.015.

DOI:10.1016/j.vetmic.2004.03.015

PMID:15172690

Abstract

Listeria ivanovii is a Gram-positive bacterial pathogen that is capable of causing abortions and stillbirths in farm animals, particularly sheep and cattle. In terms of morphological, biochemical and molecular characteristics, L. ivanovii resembles other Listeria species such as L. monocytogenes, a pathogen of both man and animals. In this study, through comparative analysis of genomic DNA from the six Listeria species, a L. ivanovii specific clone (liv22-228) containing a 946 bp insert was isolated. This clone contained the 5' ends of two divergently transcribed L. ivanovii genes and an intergenic spacer region, similar in organization to homologous regions from the L. innocua and L. monocytogenes genomes. Regions of low homology in the clone were identified by comparing to the L. innocua and L. monocytogenes genomes, and oligonucleotide primers (liv22-228F and liv22-228R) were designed. These primers amplified a 463 bp band from genomic DNA of L. ivanovii strains only, but not from other Listeria species or common bacteria. Thus, PCR employing L. ivanovii specific primers (liv22-228F and liv22-228R) provides a useful and straightforward method for rapid and precise determination of L. ivanovii.

摘要

伊氏李斯特菌是一种革兰氏阳性细菌病原体，能够导致农场动物流产和死产，尤其是绵羊和牛。在形态、生化和分子特征方面，伊氏李斯特菌与其他李斯特菌属物种相似，如人畜共患病原体单核细胞增生李斯特菌。在本研究中，通过对六种李斯特菌属物种的基因组DNA进行比较分析，分离出一个含有946 bp插入片段的伊氏李斯特菌特异性克隆（liv22-228）。该克隆包含两个反向转录的伊氏李斯特菌基因的5'端和一个基因间隔区，其组织形式与无害李斯特菌和单核细胞增生李斯特菌基因组的同源区域相似。通过与无害李斯特菌和单核细胞增生李斯特菌基因组进行比较，确定了克隆中的低同源性区域，并设计了寡核苷酸引物（liv22-228F和liv22-228R）。这些引物仅从伊氏李斯特菌菌株的基因组DNA中扩增出一条463 bp的条带，而不从其他李斯特菌属物种或常见细菌中扩增。因此，使用伊氏李斯特菌特异性引物（liv22-228F和liv22-228R）的PCR为快速、准确地鉴定伊氏李斯特菌提供了一种有用且直接的方法。