Williams Tracie L, Musser Steven M, Nordstrom Jessica L, DePaola Angelo, Monday Steven R
Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, Maryland 20740, USA.
J Clin Microbiol. 2004 Apr;42(4):1657-65. doi: 10.1128/JCM.42.4.1657-1665.2004.
The present method of characterizing Vibrio parahaemolyticus strains involves serotyping or detection methods based on assessment of the presence or absence of genes thought to be markers of an organism's pathogenicity. It is unclear whether these assays detect all pathogenic V. parahaemolyticus strains since a clear correlation between the presence of a particular gene and the organism's pathogenicity has not yet been observed. We have described a proteomics-based method to distinguish individual V. parahaemolyticus strains on the basis of their protein profiles and identified a specific protein that is characteristic of the pandemic O3:K6 strain and its clonal derivatives. In the pandemic clone of V. parahaemolyticus, a histone-like DNA-binding protein, HU-alpha, has a C-terminal amino acid sequence different from those of other strains of V. parahaemolyticus. Upon further study, it was discovered that the gene encoding this protein has a 16-kbp insert at the 3' terminus of the open reading frame for this protein. By using the protein sequence of the unique biomarker for the pandemic clone of V. parahaemolyticus, it was possible to rationally design specific PCR-based probes and assays that permit the rapid and precise identification of pandemic strains of V. parahaemolyticus.
目前对副溶血性弧菌菌株进行特征描述的方法包括血清分型或基于评估被认为是生物体致病性标志物的基因存在与否的检测方法。由于尚未观察到特定基因的存在与生物体致病性之间的明确相关性,这些检测方法是否能检测出所有致病性副溶血性弧菌菌株尚不清楚。我们描述了一种基于蛋白质组学的方法,可根据副溶血性弧菌菌株的蛋白质谱来区分各个菌株,并鉴定出一种特定蛋白质,它是大流行的O3:K6菌株及其克隆衍生物的特征。在副溶血性弧菌的大流行克隆中,一种类组蛋白DNA结合蛋白HU-α的C末端氨基酸序列与其他副溶血性弧菌菌株不同。进一步研究发现,编码该蛋白的基因在该蛋白开放阅读框的3'末端有一个16-kbp的插入片段。通过使用副溶血性弧菌大流行克隆的独特生物标志物的蛋白质序列,有可能合理设计基于PCR的特异性探针和检测方法,从而快速、准确地鉴定副溶血性弧菌的大流行菌株。