目的探讨广州地区耐多药结核分枝杆菌散在重复单元-数目可变串联重复序列(ycobacterial interspersed repetitive unit-variable number tandem repeat,MIRU-VNTR)与耐药性变迁的关系。方法对2011至2015年广州市胸科医院收治的36例耐多...目的探讨广州地区耐多药结核分枝杆菌散在重复单元-数目可变串联重复序列(ycobacterial interspersed repetitive unit-variable number tandem repeat,MIRU-VNTR)与耐药性变迁的关系。方法对2011至2015年广州市胸科医院收治的36例耐多药结核病患者在治疗期间的多次分离培养株105株(2~8株不等)进行MIRU-VNTR分型,分析在治疗期间异烟肼、乙胺丁醇、链霉素、阿米卡星、左氧氟沙星和莫西沙星等耐药性有变化者与16个MIRU-VNTR位点多态性之间的相关性。耐药表型和VNTR位点之间的相关性使用χ^(2)检验。结果61.1%(22/36)的患者(69株)发生了耐药性变迁,其中异烟肼耐药表型变迁结核菌4株(5.8%),乙胺丁醇耐药表型变迁结核菌18株(26.1%),链霉素耐药表型变迁结核菌8株(11.6%),阿米卡星耐药表型变迁结核菌8株(11.6%),左氧氟沙星耐药表型变迁结核菌21株(30.4%),莫西沙星耐药表型变迁结核菌25株(36.2%)。耐药表型变迁百分率顺序:莫西沙星>左氧氟沙星>乙胺丁醇>链霉素/阿米卡星>异烟肼,6种耐药表型均存在菌株由敏感变为耐药或耐药变为敏感的情况。耐多药结核分枝杆菌的耐药变迁表型与VNTR位点的整体关联差异无统计学意义(P>0.05),耐药变迁表型无变化与具有4种耐药变迁表型变化的VNTR位点关联差异有统计学意义(P<0.05)。乙胺丁醇耐药表型-VNTR4348、链霉素耐药表型-VNTR0595、莫西沙星耐药表型-VNTR1907和莫西沙星耐药表型-VNTR1895差异均有统计学意义(均P<0.05)。结论耐多药结核病患者病原菌耐药性变迁与MIRU-VNTR多态性存在关联,具体机制尚待进一步研究。展开更多
Background: Molecular typing allows a rapid and precise species differentiation and is essential in investigating the spread of specific genotypes and any relationship with drug resistance. Methodology: To compare the...Background: Molecular typing allows a rapid and precise species differentiation and is essential in investigating the spread of specific genotypes and any relationship with drug resistance. Methodology: To compare the discrimination power of 24-loci Mycobacteria interspersed repetitive units-variable number of tandem repeat (MIRU-VNTR) to spoligotyping in determining the circulating genotypes of Mycobacterium tuberculosis in isolates from pulmonary tuberculosis patients in Kenya, a total of 204 isolates were typed. Results: Spoligotyping identified 22 spoligo lineages;while 36(17.6%) isolates were not determined. MIRU-VNTR typing identified 12 genotypes;Kenya H37_Rv_ like, S-like that had never been reported before and which were not identified by spoligotyping were identified. Others were Uganda I and II, LAM, Beijing, TUR, EAI, Delhi/C, S and Haarlem. Only 8 (3.9%) were not defined by MIRU-VNTR. Delhi/CAS, EAI, S, S-like, LAM and Beijing had strains that showed resistance to all the five drugs tested. Two strains of EAI and 2 of S genotypes were resistant to all the five drugs tested. Beijing genotype commonly associated with drug resistance was found to be third in drug resistance (14.7%) after Delhi/CAS (28.9%) and LAM (17.6%) with the highest resistance towards isoniazid and pyrazinamide (3.9% each). MIRU-VNTR typing was more discriminative than spoligotyping;identifying 10 unique H37_Rv-like isolates designated KeniaH37_Rv_like genotype and 14 S-like genotype. Conclusion: MIRU-VNTR typing has not been reported in any other study in Kenya and its higher discrimination can help identify genotypes that cannot be determined by spoligotyping. Association of Beijing genotype drug resistance particularly isoniazid should be of concern since it may result in multidrug resistance in the patients.展开更多
目的探讨河南省结核杆菌的遗传多样性。方法使用26位点(经典24位点和其他2位点)的分支杆菌散在重复单元中数目可变串联重复序列(mycobacterium interspersed repetitive unit-variable number tandem repeat,MIRU-VNTR)和间隔区寡核苷...目的探讨河南省结核杆菌的遗传多样性。方法使用26位点(经典24位点和其他2位点)的分支杆菌散在重复单元中数目可变串联重复序列(mycobacterium interspersed repetitive unit-variable number tandem repeat,MIRU-VNTR)和间隔区寡核苷酸分型(spoligotyping)对来自2015年期间河南省不同地区的668株结核杆菌分离株进行分型分析。对结核菌spoligotype类型和耐药表型的相关性进行分析。结果 Spoligotyping分析表明668株结核杆菌被分到10个不同的基因簇中。北京家族基因型是河南地区结核杆菌中的优势菌株。北京家族基因型菌株对四种一线药全耐药以及耐多药菌株比例明显高于非北京型菌株。668株结核杆菌通过26位点VNTR被分为567个不同的基因型。26个位点中15个位点具有高度或适中的分辨能力。10个分辨能力最强位点的组合分辨力能与26位点相当。结论北京家族是河南省最流行的结核杆菌基因型。10位点VNTR和spoligotyping相结合可有效地用于河南省结核杆菌进化遗传学研究。展开更多
目的了解大理地区结核分枝杆菌基因组中的数目可变串联重复序列(variable number tandem repeat,VNTR)即MIRU(mycobacterium interspersed repetitive unit)基因多态性,探讨MIRU-VNTR位点多态性的应用价值。方法采用PCR和琼脂糖凝胶电...目的了解大理地区结核分枝杆菌基因组中的数目可变串联重复序列(variable number tandem repeat,VNTR)即MIRU(mycobacterium interspersed repetitive unit)基因多态性,探讨MIRU-VNTR位点多态性的应用价值。方法采用PCR和琼脂糖凝胶电泳检测大理地区临床分离的60株结核分枝杆菌7个VNTR位点,采用MIRU-VNTR技术进行分型,利用Hunter-Gaston指数(HGI)对各MIRU进行分辨力评价,并应用Quantity one软件和BioNumerics6.6软件进行数字化和聚类分析。结果 60株结核分枝杆菌分为5个基因群(Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ群)52个基因型。Ⅰ群占56.77%,含有29个基因型;Ⅱ群占25.00%,含有13个基因型;Ⅲ群占8.33%,含有5个基因型;Ⅳ群占6.67%,含有3个基因型;Ⅴ群占3.33%,含有2个基因型。7位点组合总分辨力为0.900,最高ETR-E位点0.735,最低ETR-C位点0.455。结论大理地区分离株结核分枝杆菌存在基因多态性,7位点MIRU-VNTR分型简便快速,分辨力高,适合用于大理地区结核分枝杆菌的基因分型检测。展开更多
耐多药结核病仍然是全球重要的公共卫生问题之一。多位点数目可变串联重复序列分析(mycobacterial interspersed repetitive unit-variable number tandem repeat typing,MIRU-VNTR)方法是一种简单快速的结核分枝杆菌基因分型方法,与其...耐多药结核病仍然是全球重要的公共卫生问题之一。多位点数目可变串联重复序列分析(mycobacterial interspersed repetitive unit-variable number tandem repeat typing,MIRU-VNTR)方法是一种简单快速的结核分枝杆菌基因分型方法,与其他分型方法结合,表现了与金标准RFLP相似的分辨力,已逐渐成为耐多药结核病分子流行病学研究中的重要工具。MIRU-VNTR主要应用于以下方面:(1)探索耐多药结核病传播机制;(2)接触者调查和爆发同源性调查;(3)区分再感染与复燃;(4)揭示结核杆菌菌种多样性和群体遗传关系;(5)与GPS结合,描述各地结核病传播风险"热点"地图;(6)预测结核分支杆菌耐药性。展开更多
目的通过对MIRU-VNTR分型技术在新疆乌鲁木齐市耐药结核病基因分型的研究,明确乌鲁木齐市12个标准VNTR分型位点的稳定性,为结核病耐药菌株演变、病源追踪、暴发调查及快速诊断等方面提供理论依据。方法采用间隔重复单元的可变数目串联...目的通过对MIRU-VNTR分型技术在新疆乌鲁木齐市耐药结核病基因分型的研究,明确乌鲁木齐市12个标准VNTR分型位点的稳定性,为结核病耐药菌株演变、病源追踪、暴发调查及快速诊断等方面提供理论依据。方法采用间隔重复单元的可变数目串联重复序列(Mycobacteral interspersed repetive units-Variable number tandem repeat,MIRU-VNTR)分型方法,选择标准12位点VNTR,对65例耐药结核分枝杆菌进行DNA检测,使用MIRU-VNTR数据分析网站进行聚类分析,并进行分辨率指数(HGI)及VNTR等位基因多态性分析。结果 12个VNTR位点等位基因多态性存在差异;除MIRU26位点分辨率较低(h=0.3381)外,其余各位点分辨率均较高(h>0.6);65株菌株可分4大群、3种基因型,成5簇,成簇率为29.231%;结论乌鲁木齐市耐药结核分枝杆菌12个标准VNTR位点等位基因多态性保持较为稳定;采用标准12位点MIRU-VNTR技术对耐药结核病进行基因分型、成簇性分析等分子流行病研究,所得结果可靠。展开更多
文摘目的探讨广州地区耐多药结核分枝杆菌散在重复单元-数目可变串联重复序列(ycobacterial interspersed repetitive unit-variable number tandem repeat,MIRU-VNTR)与耐药性变迁的关系。方法对2011至2015年广州市胸科医院收治的36例耐多药结核病患者在治疗期间的多次分离培养株105株(2~8株不等)进行MIRU-VNTR分型,分析在治疗期间异烟肼、乙胺丁醇、链霉素、阿米卡星、左氧氟沙星和莫西沙星等耐药性有变化者与16个MIRU-VNTR位点多态性之间的相关性。耐药表型和VNTR位点之间的相关性使用χ^(2)检验。结果61.1%(22/36)的患者(69株)发生了耐药性变迁,其中异烟肼耐药表型变迁结核菌4株(5.8%),乙胺丁醇耐药表型变迁结核菌18株(26.1%),链霉素耐药表型变迁结核菌8株(11.6%),阿米卡星耐药表型变迁结核菌8株(11.6%),左氧氟沙星耐药表型变迁结核菌21株(30.4%),莫西沙星耐药表型变迁结核菌25株(36.2%)。耐药表型变迁百分率顺序:莫西沙星>左氧氟沙星>乙胺丁醇>链霉素/阿米卡星>异烟肼,6种耐药表型均存在菌株由敏感变为耐药或耐药变为敏感的情况。耐多药结核分枝杆菌的耐药变迁表型与VNTR位点的整体关联差异无统计学意义(P>0.05),耐药变迁表型无变化与具有4种耐药变迁表型变化的VNTR位点关联差异有统计学意义(P<0.05)。乙胺丁醇耐药表型-VNTR4348、链霉素耐药表型-VNTR0595、莫西沙星耐药表型-VNTR1907和莫西沙星耐药表型-VNTR1895差异均有统计学意义(均P<0.05)。结论耐多药结核病患者病原菌耐药性变迁与MIRU-VNTR多态性存在关联,具体机制尚待进一步研究。
文摘Background: Molecular typing allows a rapid and precise species differentiation and is essential in investigating the spread of specific genotypes and any relationship with drug resistance. Methodology: To compare the discrimination power of 24-loci Mycobacteria interspersed repetitive units-variable number of tandem repeat (MIRU-VNTR) to spoligotyping in determining the circulating genotypes of Mycobacterium tuberculosis in isolates from pulmonary tuberculosis patients in Kenya, a total of 204 isolates were typed. Results: Spoligotyping identified 22 spoligo lineages;while 36(17.6%) isolates were not determined. MIRU-VNTR typing identified 12 genotypes;Kenya H37_Rv_ like, S-like that had never been reported before and which were not identified by spoligotyping were identified. Others were Uganda I and II, LAM, Beijing, TUR, EAI, Delhi/C, S and Haarlem. Only 8 (3.9%) were not defined by MIRU-VNTR. Delhi/CAS, EAI, S, S-like, LAM and Beijing had strains that showed resistance to all the five drugs tested. Two strains of EAI and 2 of S genotypes were resistant to all the five drugs tested. Beijing genotype commonly associated with drug resistance was found to be third in drug resistance (14.7%) after Delhi/CAS (28.9%) and LAM (17.6%) with the highest resistance towards isoniazid and pyrazinamide (3.9% each). MIRU-VNTR typing was more discriminative than spoligotyping;identifying 10 unique H37_Rv-like isolates designated KeniaH37_Rv_like genotype and 14 S-like genotype. Conclusion: MIRU-VNTR typing has not been reported in any other study in Kenya and its higher discrimination can help identify genotypes that cannot be determined by spoligotyping. Association of Beijing genotype drug resistance particularly isoniazid should be of concern since it may result in multidrug resistance in the patients.
文摘目的探讨河南省结核杆菌的遗传多样性。方法使用26位点(经典24位点和其他2位点)的分支杆菌散在重复单元中数目可变串联重复序列(mycobacterium interspersed repetitive unit-variable number tandem repeat,MIRU-VNTR)和间隔区寡核苷酸分型(spoligotyping)对来自2015年期间河南省不同地区的668株结核杆菌分离株进行分型分析。对结核菌spoligotype类型和耐药表型的相关性进行分析。结果 Spoligotyping分析表明668株结核杆菌被分到10个不同的基因簇中。北京家族基因型是河南地区结核杆菌中的优势菌株。北京家族基因型菌株对四种一线药全耐药以及耐多药菌株比例明显高于非北京型菌株。668株结核杆菌通过26位点VNTR被分为567个不同的基因型。26个位点中15个位点具有高度或适中的分辨能力。10个分辨能力最强位点的组合分辨力能与26位点相当。结论北京家族是河南省最流行的结核杆菌基因型。10位点VNTR和spoligotyping相结合可有效地用于河南省结核杆菌进化遗传学研究。
文摘目的了解大理地区结核分枝杆菌基因组中的数目可变串联重复序列(variable number tandem repeat,VNTR)即MIRU(mycobacterium interspersed repetitive unit)基因多态性,探讨MIRU-VNTR位点多态性的应用价值。方法采用PCR和琼脂糖凝胶电泳检测大理地区临床分离的60株结核分枝杆菌7个VNTR位点,采用MIRU-VNTR技术进行分型,利用Hunter-Gaston指数(HGI)对各MIRU进行分辨力评价,并应用Quantity one软件和BioNumerics6.6软件进行数字化和聚类分析。结果 60株结核分枝杆菌分为5个基因群(Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ群)52个基因型。Ⅰ群占56.77%,含有29个基因型;Ⅱ群占25.00%,含有13个基因型;Ⅲ群占8.33%,含有5个基因型;Ⅳ群占6.67%,含有3个基因型;Ⅴ群占3.33%,含有2个基因型。7位点组合总分辨力为0.900,最高ETR-E位点0.735,最低ETR-C位点0.455。结论大理地区分离株结核分枝杆菌存在基因多态性,7位点MIRU-VNTR分型简便快速,分辨力高,适合用于大理地区结核分枝杆菌的基因分型检测。
文摘耐多药结核病仍然是全球重要的公共卫生问题之一。多位点数目可变串联重复序列分析(mycobacterial interspersed repetitive unit-variable number tandem repeat typing,MIRU-VNTR)方法是一种简单快速的结核分枝杆菌基因分型方法,与其他分型方法结合,表现了与金标准RFLP相似的分辨力,已逐渐成为耐多药结核病分子流行病学研究中的重要工具。MIRU-VNTR主要应用于以下方面:(1)探索耐多药结核病传播机制;(2)接触者调查和爆发同源性调查;(3)区分再感染与复燃;(4)揭示结核杆菌菌种多样性和群体遗传关系;(5)与GPS结合,描述各地结核病传播风险"热点"地图;(6)预测结核分支杆菌耐药性。
文摘目的通过对MIRU-VNTR分型技术在新疆乌鲁木齐市耐药结核病基因分型的研究,明确乌鲁木齐市12个标准VNTR分型位点的稳定性,为结核病耐药菌株演变、病源追踪、暴发调查及快速诊断等方面提供理论依据。方法采用间隔重复单元的可变数目串联重复序列(Mycobacteral interspersed repetive units-Variable number tandem repeat,MIRU-VNTR)分型方法,选择标准12位点VNTR,对65例耐药结核分枝杆菌进行DNA检测,使用MIRU-VNTR数据分析网站进行聚类分析,并进行分辨率指数(HGI)及VNTR等位基因多态性分析。结果 12个VNTR位点等位基因多态性存在差异;除MIRU26位点分辨率较低(h=0.3381)外,其余各位点分辨率均较高(h>0.6);65株菌株可分4大群、3种基因型,成5簇,成簇率为29.231%;结论乌鲁木齐市耐药结核分枝杆菌12个标准VNTR位点等位基因多态性保持较为稳定;采用标准12位点MIRU-VNTR技术对耐药结核病进行基因分型、成簇性分析等分子流行病研究,所得结果可靠。
