目的分析福建省产超广谱β内酰胺酶(ESBLs)宋内志贺菌基因组特征,并对共存于质粒的bla_(CTX-M)耐药基因进行快速鉴定。方法收集福建省6个地区2007—2024年产ESBLs宋内志贺菌26株,微量肉汤法检测抗菌素最小抑菌浓度(MIC);利用二代和三代...目的分析福建省产超广谱β内酰胺酶(ESBLs)宋内志贺菌基因组特征,并对共存于质粒的bla_(CTX-M)耐药基因进行快速鉴定。方法收集福建省6个地区2007—2024年产ESBLs宋内志贺菌26株,微量肉汤法检测抗菌素最小抑菌浓度(MIC);利用二代和三代测序技术进行全基因组测序(WGS),预测耐药基因、质粒、插入序列(IS)和bla_(CTX-M)基因环境,Roary构建聚类树,cgSNP(core genome single nucleotide polymorphism)分析菌株的全球进化地位。S1核酸内切酶脉冲场凝胶电泳(S1-PFGE)、微滴式数字PCR(ddPCR)及质粒接合转移试验确认bla_(CTX-M)在质粒上的定位和水平传播。结果26株产ESBLs宋内志贺菌均为多重耐药菌,全部对头孢噻肟耐药,有6株对阿奇霉素耐药。WGS分析显示,26株菌共携带17种耐药基因、9种质粒复制子、15种插入序列和1种转座子Tn7。bla_(CTX-M-14)、bla_(CTX-M-15)和bla_(CTX-M-64)、ermB和mphA的阳性率分别为61.54%、30.77%、7.69%、11.54%和11.54%,bla_(CTX-M)基因环境可分为4种类型。Roary分析可将26株菌分为4个簇;cgSNP显示,它们均归属于独特的基因型3.7.6中国分支。除1株bla_(CTX-M-14)位于染色体外,所有bla_(CTX-M)分布于3种类型的质粒上,S1-PFGE和ddPCR鉴定结果与测序结果一致。结论福建产ESBLs宋内志贺菌基因组携带大量耐药因子,S1-PFGE和ddPCR可成为检测质粒携带耐药基因的有效工具。展开更多
目的了解汕头市水产品非O1/O139群霍乱弧菌的耐药基因、毒力基因、序列位点信息,分析不同菌株间遗传关系。方法收集分离自水产品的霍乱弧菌41株,对菌株进行全基因组序列测定,利用生物信息学工具预测菌株的耐药基因和毒力基因情况,结合...目的了解汕头市水产品非O1/O139群霍乱弧菌的耐药基因、毒力基因、序列位点信息,分析不同菌株间遗传关系。方法收集分离自水产品的霍乱弧菌41株,对菌株进行全基因组序列测定,利用生物信息学工具预测菌株的耐药基因和毒力基因情况,结合美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库,进行多位点序列分型分析(multilocus sequence typing,MLST)和核心基因组单核苷酸多态性分析(core gene single nucleotide polymorphism,cgSNP)。结果41株霍乱弧菌共检测到34个抗生素耐药基因,携带率最高的是dfrA31(46.3%),部分菌株携带能水解所有β-内酰胺酶的blaNDM-1基因。毒力基因携带率最高的是rtxA(100.0%)和hlyA(97.6%),所有病原菌株均不携带ctxA和ctxB,致病岛VPI-1/VPI-2携带率分别为31.7%/70.7%,T6SS和第7次大流行特征基因岛VSP-1/VSP-2的检出率都是4.9%。MLST和cgSNP分析显示,共有30种不同的序列型(sequencetypes,STs),其中20种为新分配的STs。结论汕头市水产品中非O1/O139群霍乱弧菌携带多种毒力基因和耐药基因,其中霍乱弧菌携带blaNDM-1基因为国内首次报道。不同物种来源的菌株遗传多样性高,发现大量新STs型。展开更多
Objective To construct the detection method of vibrio cholerae with fluorescence quantitative PCR and evaluate its superiority.Methods A diagnostic kit(FQ-PCR) was developed according to the common sequence of NhaA ge...Objective To construct the detection method of vibrio cholerae with fluorescence quantitative PCR and evaluate its superiority.Methods A diagnostic kit(FQ-PCR) was developed according to the common sequence of NhaA gene in vibrio cholerae.The sensitivity and speciality in detecting of vibrio cholerae in infectious water,sea food and winter-endurance specimen and clinical samples were compared with FQ-PCR kit.Results In the detection of clinical samples,FQ-PCR and regular detection ways have the same sensitivity and speciality.While in the detection of infectious water,sea food and winter-endurance specimen,FQ-PCR has superiority to regular ways.Conclusion The construction of FQ-PCR method eliminates PCR cross-contamination which cause false positive,and real-time detection ensure accurate quantity.So FQ-PCR can be used to the detection of vibrio cholerae in clinical and sanitary laboratory.展开更多
文摘目的分析福建省产超广谱β内酰胺酶(ESBLs)宋内志贺菌基因组特征,并对共存于质粒的bla_(CTX-M)耐药基因进行快速鉴定。方法收集福建省6个地区2007—2024年产ESBLs宋内志贺菌26株,微量肉汤法检测抗菌素最小抑菌浓度(MIC);利用二代和三代测序技术进行全基因组测序(WGS),预测耐药基因、质粒、插入序列(IS)和bla_(CTX-M)基因环境,Roary构建聚类树,cgSNP(core genome single nucleotide polymorphism)分析菌株的全球进化地位。S1核酸内切酶脉冲场凝胶电泳(S1-PFGE)、微滴式数字PCR(ddPCR)及质粒接合转移试验确认bla_(CTX-M)在质粒上的定位和水平传播。结果26株产ESBLs宋内志贺菌均为多重耐药菌,全部对头孢噻肟耐药,有6株对阿奇霉素耐药。WGS分析显示,26株菌共携带17种耐药基因、9种质粒复制子、15种插入序列和1种转座子Tn7。bla_(CTX-M-14)、bla_(CTX-M-15)和bla_(CTX-M-64)、ermB和mphA的阳性率分别为61.54%、30.77%、7.69%、11.54%和11.54%,bla_(CTX-M)基因环境可分为4种类型。Roary分析可将26株菌分为4个簇;cgSNP显示,它们均归属于独特的基因型3.7.6中国分支。除1株bla_(CTX-M-14)位于染色体外,所有bla_(CTX-M)分布于3种类型的质粒上,S1-PFGE和ddPCR鉴定结果与测序结果一致。结论福建产ESBLs宋内志贺菌基因组携带大量耐药因子,S1-PFGE和ddPCR可成为检测质粒携带耐药基因的有效工具。
文摘目的了解汕头市水产品非O1/O139群霍乱弧菌的耐药基因、毒力基因、序列位点信息,分析不同菌株间遗传关系。方法收集分离自水产品的霍乱弧菌41株,对菌株进行全基因组序列测定,利用生物信息学工具预测菌株的耐药基因和毒力基因情况,结合美国国家生物技术信息中心(National Center for Biotechnology Information,NCBI)数据库,进行多位点序列分型分析(multilocus sequence typing,MLST)和核心基因组单核苷酸多态性分析(core gene single nucleotide polymorphism,cgSNP)。结果41株霍乱弧菌共检测到34个抗生素耐药基因,携带率最高的是dfrA31(46.3%),部分菌株携带能水解所有β-内酰胺酶的blaNDM-1基因。毒力基因携带率最高的是rtxA(100.0%)和hlyA(97.6%),所有病原菌株均不携带ctxA和ctxB,致病岛VPI-1/VPI-2携带率分别为31.7%/70.7%,T6SS和第7次大流行特征基因岛VSP-1/VSP-2的检出率都是4.9%。MLST和cgSNP分析显示,共有30种不同的序列型(sequencetypes,STs),其中20种为新分配的STs。结论汕头市水产品中非O1/O139群霍乱弧菌携带多种毒力基因和耐药基因,其中霍乱弧菌携带blaNDM-1基因为国内首次报道。不同物种来源的菌株遗传多样性高,发现大量新STs型。
文摘Objective To construct the detection method of vibrio cholerae with fluorescence quantitative PCR and evaluate its superiority.Methods A diagnostic kit(FQ-PCR) was developed according to the common sequence of NhaA gene in vibrio cholerae.The sensitivity and speciality in detecting of vibrio cholerae in infectious water,sea food and winter-endurance specimen and clinical samples were compared with FQ-PCR kit.Results In the detection of clinical samples,FQ-PCR and regular detection ways have the same sensitivity and speciality.While in the detection of infectious water,sea food and winter-endurance specimen,FQ-PCR has superiority to regular ways.Conclusion The construction of FQ-PCR method eliminates PCR cross-contamination which cause false positive,and real-time detection ensure accurate quantity.So FQ-PCR can be used to the detection of vibrio cholerae in clinical and sanitary laboratory.
