Phenotype and genotypic characteristics of Vibrio parahaemolyticus strains isolated from diarrhea patients in Liaoning province
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摘要: 目的 了解辽宁地区副溶血弧菌腹泻病例分离株的表型及基因型特征。方法 对2012-2013年辽宁地区101株副溶血弧菌临床分离株进行血清型分型;采用微量肉汤法,进行16种抗菌药物的药敏试验;通过PCR方法对其进行tlh、tdh和trh毒力基因检测;采用脉冲场凝胶电泳(PFGE)方法对菌株进行基因分型。结果 101株副溶血弧菌分属于25种血清型,其中O3:K6为优势血清型,占37.62%(38/101);对磺胺甲恶唑、链霉素、氨苄西林产生耐药;对环丙沙星、强力霉素、头孢吡肟、甲氧苄啶、亚胺培南、头孢曲松钠等均具有较高敏感性;全部菌株tlh基因阳性;tdh存在于大部分菌株中,所占比例为84.16%(85/101);全部菌株trh基因阴性;O3:K6型菌株tdh基因阳性率100%,明显高于非O3:K6型菌株(76.19%,77/101)(P=0.00081)。101株副溶血弧菌分成63种型别,38株O3:K6型菌株分成19种型别,带型分散,无明显聚集性。结论 辽宁省感染性腹泻病例副溶血弧菌分离株以O3:K6型为主;毒力基因tlh和tdh携带率高,且O3:K6型临床分离株毒力更强;副溶血弧菌对多数抗生素均具有较高敏感性;PFGE结果提示辽宁地区流行的副溶血弧菌存在多克隆来源。Abstract: Objective To examine the phenotype and gene characteristics of Vibrio parahaemolyticus(VPS)strains isolated from diarrhea patients in Liaoning province from 2012 to 2013.Methods A total of 101 VPS strains isolated from diarrhea patients in 2012-2013 were sreotyped.All the strains were tested for drug sensitivity with microdilution broth method.PCR was applied to detect three virulence genes tlh,tdh,and trh.Molecular typing on VPS isolates was completed with pulsed-field gel electrophoresis(PFGE)and the patterns were clustered by BioNumerics software.Results The 101 isolates were belonged to 25 serotypes,with a dominant serotype of O3:K6(37.62%,38/101).The strains isolated from the diarrhea patients were resistant to sulfamethoxazole,streptomycin,and ampicillin,while with high sensitivity to ciprofloxacin,doxycycline,cefepime,trimethoprim,imipenem,and ceftriaxone.Virulence gene detection showed that all the strains were positive to tlh and the most of the strains were positivie to tdh(84.16%,85/101),but all the strains were negative to trh.The positive rate of tdh among O3:K6 strains(100%)was higher than that among non-O3:K6 strains(76.19%,77/101)(P=0.000 81).The 101 isolates were discriminated into 63 different PFGE patterns,while 38 O3:K6 strains into 19 patterns without cluster characteristics.Conclusion Vibrio parahaemolyticus strains isolated from diarrheal patients in Liaoning province from 2012 to 2013 were dominated by O3:K6 serotype,with stronger virulence and high positive rate of tlh and tdh gene.Vibrio parahaemolyticus strains showed high sensitivity to most antibiotics.The prevalent Vibrio parahaemolyticus strains in Liaoning province present various clonal origins.
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