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鉴定一株携带 ail 基因的小肠结肠炎耶尔森氏菌生物型 1A 菌株。

Characterization of a Yersinia enterocolitica biotype 1A strain harbouring an ail gene.

机构信息

Department of Biological Safety, Federal Institute for Risk Assessment, Berlin, Germany.

出版信息

J Appl Microbiol. 2011 Oct;111(4):997-1005. doi: 10.1111/j.1365-2672.2011.05112.x. Epub 2011 Aug 11.

Abstract

AIMS

The chromosomal ail gene (attachment and invasion locus) is commonly used as target gene for the detection of pathogenic Y. enterocolitica strains in food testing. The ail PCR does not detect strains of biotype 1A (BT1A), which are regarded as non-pathogenic because BT1A strains lack the virulence plasmid and chromosomally encoded virulence genes. In some recent reports, however, BT1A strains were discovered that harboured the ail gene. We isolated an ail-positive strain and characterized this strain with phenotypic and genotypic methods to study its possible relation to pathogenic Y. enterocolitica strains.

METHODS AND RESULTS

The ail region of the BT1A strain was sequenced and compared with the corresponding region of nonpathogenic BT1A strains and pathogenic strains. Pulsed field gel electrophoresis (PFGE) analysis was applied revealing no similarity of the PFGE pattern of this strain to the patterns of pathogenic strains. Virulence-gene-based PCR analyses showed the strain to be positive for ystB, but negative for virulence genes ystA, virF and yadA. Whole-cell MALDI-TOF MS combined with a shrinkage discriminant analysis approach was applied and clearly classified the ail-positive biotype 1A strain within the cluster of BT1A strains.

CONCLUSIONS

PCR detection of ail sequences in food matrices should be followed by the isolation of the responsible strain and its characterization using phenotypic or genotypic methods.

SIGNIFICANCE AND IMPACT OF THE STUDY

The ail gene may be present in Y. enterocolitica BT1A strains, which are commonly considered as nonpathogenic. Efficient methods such as PCR typing of other virulence genes or rapid MALDI-TOF MS-based bacterial profiling allow a more comprehensive assessment of the pathogenicity potential of Yersinia strains.

摘要

目的

染色体 ail 基因(附着和入侵基因座)通常被用作食品检测中致病性 Y. enterocolitica 菌株检测的靶基因。ail PCR 无法检测生物型 1A(BT1A)菌株,因为 BT1A 菌株缺乏毒力质粒和染色体编码的毒力基因,被认为是非致病性的。然而,在最近的一些报告中,发现了携带 ail 基因的 BT1A 菌株。我们分离了一株 ail 阳性菌株,并采用表型和基因型方法对其进行了特征描述,以研究其与致病性 Y. enterocolitica 菌株的可能关系。

方法和结果

对 BT1A 菌株的 ail 区域进行测序,并与非致病性 BT1A 菌株和致病性菌株的相应区域进行比较。应用脉冲场凝胶电泳(PFGE)分析显示,该菌株的 PFGE 模式与致病性菌株的模式没有相似性。基于毒力基因的 PCR 分析显示,该菌株 ystB 基因呈阳性,但 ystA、virF 和 yadA 等毒力基因呈阴性。应用全细胞 MALDI-TOF MS 结合收缩判别分析方法,将 ail 阳性的生物型 1A 菌株明确分类为 BT1A 菌株群内的一个分支。

结论

在食品基质中使用 ail 序列 PCR 检测后,应通过分离负责菌株并使用表型或基因型方法对其进行特征描述。

研究的意义和影响

ail 基因可能存在于通常被认为是非致病性的 Y. enterocolitica BT1A 菌株中。其他毒力基因的 PCR 分型或快速 MALDI-TOF MS 基于的细菌分析等高效方法可以更全面地评估 Yersinia 菌株的致病性潜力。

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