Nilsson William B, Turner Jeffrey W
Environmental and Fisheries Science Division, Northwest Fisheries Science Center, National Marine Fisheries Service, NOAA, 2725 Montlake Blvd. E., Seattle, WA 98112, USA.
Environmental and Fisheries Science Division, Northwest Fisheries Science Center, National Marine Fisheries Service, NOAA, 2725 Montlake Blvd. E., Seattle, WA 98112, USA.
J Microbiol Methods. 2016 Jul;126:1-7. doi: 10.1016/j.mimet.2016.04.007. Epub 2016 Apr 16.
Vibrio parahaemolyticus is a leading cause of bacterial food-related illness associated with the consumption of undercooked seafood. Only a small subset of strains is pathogenic. Most clinical strains encode for the thermostable direct hemolysin (TDH) and/or the TDH-related hemolysin (TRH). In this work, we amplify and sequence the trh gene from over 80 trh+strains of this bacterium and identify thirteen genetically distinct alleles, most of which have not been deposited in GenBank previously. Sequence data was used to design new primers for more reliable detection of trh by endpoint PCR. We also designed a new quantitative PCR assay to target a more conserved gene that is genetically-linked to trh. This gene, ureR, encodes the transcriptional regulator for the urease gene cluster immediately upstream of trh. We propose that this ureR assay can be a useful screening tool as a surrogate for direct detection of trh that circumvents challenges associated with trh sequence variation.
副溶血性弧菌是与食用未煮熟海鲜相关的细菌性食源性疾病的主要病因。只有一小部分菌株具有致病性。大多数临床菌株编码耐热直接溶血素（TDH）和/或TDH相关溶血素（TRH）。在这项工作中，我们对该细菌的80多个trh+菌株的trh基因进行了扩增和测序，鉴定出13个遗传上不同的等位基因，其中大多数以前未存入GenBank。序列数据用于设计新引物，以便通过终点PCR更可靠地检测trh。我们还设计了一种新的定量PCR检测方法，以靶向一个与trh基因连锁的更保守的基因。这个基因ureR编码紧接在trh上游的脲酶基因簇的转录调节因子。我们认为，这种ureR检测方法可以作为一种有用的筛选工具，作为直接检测trh的替代方法，规避与trh序列变异相关的挑战。
