目的通过对碳青霉烯类耐药鲍氏不动杆菌进行3种分子流行病学调查方法的对比研究,了解脉冲场凝胶电泳(PFGE)、多位点序列分析(MLST)和多位点可变数目串联重复序列分析(MLVA)3种方法在对碳青霉烯类耐药鲍氏不动杆菌复合体的流行病学调查...目的通过对碳青霉烯类耐药鲍氏不动杆菌进行3种分子流行病学调查方法的对比研究,了解脉冲场凝胶电泳(PFGE)、多位点序列分析(MLST)和多位点可变数目串联重复序列分析(MLVA)3种方法在对碳青霉烯类耐药鲍氏不动杆菌复合体的流行病学调查中的优缺点和分辨率。方法收集某三级医院2011-2012年分离自痰标本的碳青霉烯类耐药鲍氏不动杆菌复合体212株,采用PFGE、MLST和MLVA方法对其进行分子流行病学分析,运用Simpson相关系数(Simpson's index of diversity)和Wallace coefficient,对3种方法的分辨率和同质性进行比较。结果 212株碳青霉烯类耐药鲍氏不动杆菌复合体,其中149株获得PFGE可分析结果,200株获得MLST可分析结果,205株获得MLVA可分析性结果;PFGE结果显示,B型菌株为优势菌株(102株,占68.46%)。MLST分型结果显示,ST195占优势(60株,占30%),其次为ST208(43株,占21.5%)和ST643(33株,占16.5%)。MLVA分型结果显示,D型菌株占优势(151株,占73.66%),其次为N型菌株(8株,占3.9%),K、M、T、W型菌株并列第三(均为4株,各占1.95%)。优势菌株主要分布在ICU。PFGE、MLST、MLVA的Simpson相关系数(D值)分别为0.46、0.76、0.94。MLVA与MLST的Wallace coefficient值略高为0.76。结论 PFGE是区域内分析克隆时间和空间分布的重要工具,本研究中院内流行的碳青霉烯类耐药鲍氏不动杆菌主要来自于ICU;MLST可全球范围内开展抗菌药物抵抗菌株,以及新变异菌株的流行病学研究;MLVA数据可用于不同实验室之间的相互交换、整合与分析。MLVA的分辨率较高,MLVA与MLST的同质性较高。展开更多
目的了解不同血清群致病性钩体的脉冲场凝胶电泳(pulsed-field gel electrophoresis,PFGE)带型特征。方法应用PFGE方法对65株不同血清群致病性钩体国际、国内参考菌株和疫苗株进行分型,并以此聚类分析,同时与菌株的多位点序列分型(multi...目的了解不同血清群致病性钩体的脉冲场凝胶电泳(pulsed-field gel electrophoresis,PFGE)带型特征。方法应用PFGE方法对65株不同血清群致病性钩体国际、国内参考菌株和疫苗株进行分型,并以此聚类分析,同时与菌株的多位点序列分型(multilocus sequence typing,MLST)、多位点串联重复序列数变化分析(multilocus VNTR analysis,MLVA)分型结果进行比较研究。结果不同血清群致病性钩体基因组DNA经NotⅠ酶切电泳后,获得了较高清晰度酶切图谱,各条带分离良好。65株钩体菌株分为61种PFGE型别。相同的血清群的钩体菌株的PFGE型别不尽相同。比较分析结果显示尽管PFGE、MLST和MLVA之间的分型结果不尽相同,但都具有较高的分辨率。聚类分析显示这些菌株可形成11个大小不一进化簇,呈现多点分散进化特征。结论获得致病性钩体分子分型的第一手资料,为后续钩体分子溯源及流行病学研究提供科学指导。展开更多
目的鉴定云南省怒江傈僳族自治州首起聚集性感染布鲁氏菌的种型和基因型特征。方法收集2023年4月怒江傈僳族自治州兰坪县报告的7例疑似聚集性布鲁氏菌病(简称布病)病例信息,采集血样分离培养菌株;采用血清学方法进行布病诊断;采用BCSP31...目的鉴定云南省怒江傈僳族自治州首起聚集性感染布鲁氏菌的种型和基因型特征。方法收集2023年4月怒江傈僳族自治州兰坪县报告的7例疑似聚集性布鲁氏菌病(简称布病)病例信息,采集血样分离培养菌株;采用血清学方法进行布病诊断;采用BCSP31-PCR和AMOS-PCR对病原菌进行种属和种型鉴定;采用多位点可变数目串联重复序列分析(multiple locus variable-number tandem repeat analysis,MLVA)技术对菌株进行基因分型,并分析菌株与我国其他地区菌株之间的亲缘关系。结果7例病例均被诊断为布病,为同村村民,日常接触羊群。共分离4株疑似布鲁氏菌菌株,经BCSP31-PCR和AMOS-PCR鉴定为羊种布鲁氏菌;4株分离菌株的MLVA-16基因型一致,MLVA-8基因型为42型,MLVA-11基因型为180型,属于东地中海谱系;与我国其他地区分离菌株的MLVA-16比较,4株分离菌株形成独立分支。结论云南省怒江傈僳族自治州首起聚集性感染布鲁氏菌的病原菌为羊种布鲁氏菌,4株分离菌株呈现独特的MLVA基因型。展开更多
Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not ...Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not been fully elucidated.To characterize the infecting strains isolated from humans,multiple-locus variable-number tandem repeats analysis(MLVA)and whole-genome single-nucleotide polymorphism(SNP)-based approaches were employed.Methods:Strains were isolated from two males blood cultures that were confirmed Brucella m elitensis positive following biotyping and MLVA.Genomic DNA was extracted from these two strains,and whole-genome sequencing was performed.Next,SNP-based phylogenetic analysis was performed to compare the two strains to 94 B.melitensis strains(complete genome and draft genome)retrieved from online databases.Results:The two Brucella isolates were identified as B.melitensis biovar 3(QH2019001 and QH2019005)following conventional biotyping and were found to have differences in their variable number tandem repeats(VNTRs)using MLVA-16.Phylogenetic examination assigned the 96 strains to five genotype groups,with QH2019001 and QH2019005 assigned to the same group,but different subgroups.Moreover,the QH2019005 strain was assigned to a new subgenotype,llj,within genotype II.These findings were then combined to determine the geographic origin of the two Brucella strains.Conclusions:Utilizing a whole-genome SNP-based approach enabled differences between the two B.m elitensis strains to be more clearly resolved,and facilitated the elucidation of their different evolutionary histories.This approach also revealed that QH2019005 is a member of a new subgenotype(Hj)with an ancient origin in the eastern Mediterranean Sea.展开更多
文摘目的掌握江苏省2018年人感染布鲁氏菌主要流行株的种型和基因型。方法运用普通PCR及AMOS多重PCR确认分离株的生物种型;采用多位点序列分型(Multilocus sequence analysis,MLSA)和多位点串联重复序列分析(Multiple-locus variable number tandem repeat analysis,MLVA)鉴定基因型,并与国内外流行株进行聚类分析。结果2018年共分离到56株布鲁氏菌,MLSA分型显示一株菌为猪种布鲁氏菌ST(Sequence type)17型,其他均为羊种布鲁氏菌ST8型。MLVA将56株菌分为47个基因亚型(46个羊种,1个猪种),聚类显示羊种布鲁氏菌全部为“东地中海簇”。结论2018年江苏省人感染布病主要为“东地中海簇”的ST8型羊种布鲁氏菌,并首次发现一例人感染ST17型猪种布鲁氏菌。
文摘目的通过对碳青霉烯类耐药鲍氏不动杆菌进行3种分子流行病学调查方法的对比研究,了解脉冲场凝胶电泳(PFGE)、多位点序列分析(MLST)和多位点可变数目串联重复序列分析(MLVA)3种方法在对碳青霉烯类耐药鲍氏不动杆菌复合体的流行病学调查中的优缺点和分辨率。方法收集某三级医院2011-2012年分离自痰标本的碳青霉烯类耐药鲍氏不动杆菌复合体212株,采用PFGE、MLST和MLVA方法对其进行分子流行病学分析,运用Simpson相关系数(Simpson's index of diversity)和Wallace coefficient,对3种方法的分辨率和同质性进行比较。结果 212株碳青霉烯类耐药鲍氏不动杆菌复合体,其中149株获得PFGE可分析结果,200株获得MLST可分析结果,205株获得MLVA可分析性结果;PFGE结果显示,B型菌株为优势菌株(102株,占68.46%)。MLST分型结果显示,ST195占优势(60株,占30%),其次为ST208(43株,占21.5%)和ST643(33株,占16.5%)。MLVA分型结果显示,D型菌株占优势(151株,占73.66%),其次为N型菌株(8株,占3.9%),K、M、T、W型菌株并列第三(均为4株,各占1.95%)。优势菌株主要分布在ICU。PFGE、MLST、MLVA的Simpson相关系数(D值)分别为0.46、0.76、0.94。MLVA与MLST的Wallace coefficient值略高为0.76。结论 PFGE是区域内分析克隆时间和空间分布的重要工具,本研究中院内流行的碳青霉烯类耐药鲍氏不动杆菌主要来自于ICU;MLST可全球范围内开展抗菌药物抵抗菌株,以及新变异菌株的流行病学研究;MLVA数据可用于不同实验室之间的相互交换、整合与分析。MLVA的分辨率较高,MLVA与MLST的同质性较高。
文摘目的鉴定云南省怒江傈僳族自治州首起聚集性感染布鲁氏菌的种型和基因型特征。方法收集2023年4月怒江傈僳族自治州兰坪县报告的7例疑似聚集性布鲁氏菌病(简称布病)病例信息,采集血样分离培养菌株;采用血清学方法进行布病诊断;采用BCSP31-PCR和AMOS-PCR对病原菌进行种属和种型鉴定;采用多位点可变数目串联重复序列分析(multiple locus variable-number tandem repeat analysis,MLVA)技术对菌株进行基因分型,并分析菌株与我国其他地区菌株之间的亲缘关系。结果7例病例均被诊断为布病,为同村村民,日常接触羊群。共分离4株疑似布鲁氏菌菌株,经BCSP31-PCR和AMOS-PCR鉴定为羊种布鲁氏菌;4株分离菌株的MLVA-16基因型一致,MLVA-8基因型为42型,MLVA-11基因型为180型,属于东地中海谱系;与我国其他地区分离菌株的MLVA-16比较,4株分离菌株形成独立分支。结论云南省怒江傈僳族自治州首起聚集性感染布鲁氏菌的病原菌为羊种布鲁氏菌,4株分离菌株呈现独特的MLVA基因型。
基金supported by National Key R&D Program of China(Grant No.2020YFA0907101)Major Infectious Diseases such as AIDS and Viral Hepatitis Prevention and Control Technology Major Projects(2018ZX10201002)the National Natural Scientific Foundation of China(81860588).
文摘Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not been fully elucidated.To characterize the infecting strains isolated from humans,multiple-locus variable-number tandem repeats analysis(MLVA)and whole-genome single-nucleotide polymorphism(SNP)-based approaches were employed.Methods:Strains were isolated from two males blood cultures that were confirmed Brucella m elitensis positive following biotyping and MLVA.Genomic DNA was extracted from these two strains,and whole-genome sequencing was performed.Next,SNP-based phylogenetic analysis was performed to compare the two strains to 94 B.melitensis strains(complete genome and draft genome)retrieved from online databases.Results:The two Brucella isolates were identified as B.melitensis biovar 3(QH2019001 and QH2019005)following conventional biotyping and were found to have differences in their variable number tandem repeats(VNTRs)using MLVA-16.Phylogenetic examination assigned the 96 strains to five genotype groups,with QH2019001 and QH2019005 assigned to the same group,but different subgroups.Moreover,the QH2019005 strain was assigned to a new subgenotype,llj,within genotype II.These findings were then combined to determine the geographic origin of the two Brucella strains.Conclusions:Utilizing a whole-genome SNP-based approach enabled differences between the two B.m elitensis strains to be more clearly resolved,and facilitated the elucidation of their different evolutionary histories.This approach also revealed that QH2019005 is a member of a new subgenotype(Hj)with an ancient origin in the eastern Mediterranean Sea.
