Haendiges Julie, Jones Jessica, Myers Robert A, Mitchell Clifford S, Butler Erin, Toro Magaly, Gonzalez-Escalona Narjol
Department of Health and Mental Hygiene, Baltimore, Maryland, USA.
Division of Seafood Science and Technology, Gulf Coast Seafood Laboratory, Food and Drug Administration, Dauphin Island, Alabama, USA.
Appl Environ Microbiol. 2016 May 16;82(11):3208-3216. doi: 10.1128/AEM.00096-16. Print 2016 Jun 1.
In the summer of 2010, Vibrio parahaemolyticus caused an outbreak in Maryland linked to the consumption of oysters. Strains isolated from both stool and oyster samples were indistinguishable by pulsed-field gel electrophoresis (PFGE). However, the oysters contained other potentially pathogenic V. parahaemolyticus strains exhibiting different PFGE patterns. In order to assess the identity, genetic makeup, relatedness, and potential pathogenicity of the V. parahaemolyticus strains, we sequenced 11 such strains (2 clinical strains and 9 oyster strains). We analyzed these genomes by in silico multilocus sequence typing (MLST) and determined their phylogeny using a whole-genome MLST (wgMLST) analysis. Our in silico MLST analysis identified six different sequence types (STs) (ST8, ST676, ST810, ST811, ST34, and ST768), with both of the clinical and four of the oyster strains being identified as belonging to ST8. Using wgMLST, we showed that the ST8 strains from clinical and oyster samples were nearly indistinguishable and belonged to the same outbreak, confirming that local oysters were the source of the infections. The remaining oyster strains were genetically diverse, differing in >3,000 loci from the Maryland ST8 strains. eBURST analysis comparing these strains with strains of other STs available at the V. parahaemolyticus MLST website showed that the Maryland ST8 strains belonged to a clonal complex endemic to Asia. This indicates that the ST8 isolates from clinical and oyster sources were likely not endemic to Maryland. Finally, this study demonstrates the utility of whole-genome sequencing (WGS) and associated analyses for source-tracking investigations.
Vibrio parahaemolyticus is an important foodborne pathogen and the leading cause of bacterial infections in the United States associated with the consumption of seafood. In the summer of 2010, Vibrio parahaemolyticus caused an outbreak in Maryland linked to oyster consumption. Strains isolated from stool and oyster samples were indistinguishable by pulsed-field gel electrophoresis (PFGE). The oysters also contained other potentially pathogenic V. parahaemolyticus strains with different PFGE patterns. Since their identity, genetic makeup, relatedness, and potential pathogenicity were unknown, their genomes were determined by using next-generation sequencing. Whole-genome sequencing (WGS) analysis by whole-genome multilocus sequence typing (wgMLST) allowed (i) identification of clinical and oyster strains with matching PFGE profiles as belonging to ST8, (ii) determination of oyster strain diversity, and (iii) identification of the clinical strains as belonging to a clonal complex (CC) described only in Asia. Finally, WGS and associated analyses demonstrated their utility for trace-back investigations.
2010年夏季,副溶血性弧菌在马里兰州引发了一起与食用牡蛎有关的疫情。从粪便和牡蛎样本中分离出的菌株通过脉冲场凝胶电泳(PFGE)无法区分。然而,这些牡蛎中还含有其他具有不同PFGE模式的潜在致病性副溶血性弧菌菌株。为了评估副溶血性弧菌菌株的身份、基因组成、亲缘关系和潜在致病性,我们对11株此类菌株(2株临床菌株和9株牡蛎菌株)进行了测序。我们通过计算机多位点序列分型(MLST)分析这些基因组,并使用全基因组MLST(wgMLST)分析确定它们的系统发育。我们的计算机MLST分析确定了六种不同的序列类型(STs)(ST8、ST676、ST810、ST811、ST34和ST768),其中临床菌株和四株牡蛎菌株被鉴定为属于ST8。使用wgMLST,我们表明来自临床和牡蛎样本的ST8菌株几乎无法区分,属于同一疫情爆发,证实当地牡蛎是感染源。其余的牡蛎菌株在基因上具有多样性,与马里兰州的ST8菌株在3000多个位点上存在差异。将这些菌株与副溶血性弧菌MLST网站上其他STs的菌株进行eBURST分析表明,马里兰州的ST8菌株属于亚洲特有的一个克隆复合体。这表明来自临床和牡蛎来源的ST8分离株可能并非马里兰州本地的。最后,本研究证明了全基因组测序(WGS)及相关分析在溯源调查中的实用性。
副溶血性弧菌是一种重要的食源性病原体,是美国与食用海鲜相关的细菌感染的主要原因。2010年夏季,副溶血性弧菌在马里兰州引发了一起与食用牡蛎有关的疫情。从粪便和牡蛎样本中分离出的菌株通过脉冲场凝胶电泳(PFGE)无法区分。这些牡蛎中还含有其他具有不同PFGE模式的潜在致病性副溶血性弧菌菌株。由于它们的身份、基因组成、亲缘关系和潜在致病性未知,因此使用下一代测序技术确定了它们的基因组。通过全基因组多位点序列分型(wgMLST)进行全基因组测序(WGS)分析能够(i)将PFGE图谱匹配的临床和牡蛎菌株鉴定为属于ST8,(ii)确定牡蛎菌株的多样性,以及(iii)将临床菌株鉴定为属于仅在亚洲描述过的一个克隆复合体(CC)。最后,WGS及相关分析证明了它们在追溯调查中的实用性。