华南地区380株结核分枝杆菌异烟肼耐药基因突变情况分析

目的探讨华南地区异烟肼耐药结核分枝杆菌临床分离株katG、mabA-inhA、oxyR-ahpC、kasA和ndh基因的突变特点。方法在广州地区分枝杆菌菌株库中, 选择异烟肼耐药性测定结果明确的380株来自华南地区结核病患者的结核分枝杆菌临床分离株, 其中异烟肼耐药株236株和异烟肼敏感株144株, 对380株结核分枝杆菌临床分离株的5种异烟肼耐药基因片段进行序列测定分析。描述性分析各基因突变出现的频率。结果在INH耐药株和敏感株中分别发现26个位点29种突变类型、16个位点17种突变类型, 均以katG基因315AGC(S)→ACC(T)、mabA-inhA基因-15T→C最为常见, 发生率分别为48.73%(115/236)、17.37%(41/236)和9.72%(14/144)、9.72%(14/144);在katG、mabA-inhA、oxyR-ahpC、kasA和ndh中分别发现3个位点5种突变、5个位点5种突变、11个位点13种突变、6个位点6种突变和11个位点11种突变, 在INH耐药株和敏感株中5个基因突变的发生率分别为52.12%(123/236)、22.46%(5...     

基因芯片技术诊断耐多药结核病的临床应用研究

摘要： 目的 探讨基因芯片法检测耐多药结核分枝杆菌临床分离株耐药相关基因 rpoB、katG 和 inhA 突变的特点, 评估其临床应用价值。 方法 选取 2013—2014 年石家庄地区耐多药结核分枝杆菌临床分离株 76 株, 采用基因芯片法检测 rpoB、katG 及 inhA 基因耐药突变位点及频率, 以比例法药敏结果为金标准, 评价基因芯片法检测菌株耐药的准确度、灵敏度和特异度, Kappa 检验评价基因芯片法和比例法药敏结果的一致性。 结果 在 76 株石家庄地区耐多药结核分枝杆菌临床分离株中, 69 株检测到 katG/inhA 基因发生单一或联合突变, 其中 katG 基因优势突变位点为 315, 占 89.9%（62/69）, 5.8%（4/69）存在 inhA -15（C→T）突变, 4.3%（3/69）发生 katG 315 位点和 inhA 启动子区域的联合突变。 73 株检测到 rpoB 基因发生突变, 优势突变位点为 531, 占 64.4%（47/73）; 其次是 526, 突变率为 15.1%（11/73）; 12.3%（9/73）发生 516 位点突变; 1.4%（1/73）发生 513 位点突变; 1.4%（1/73）发生 533 位点突变; 另有 5.5%（4/73）发生多位点的联合突变。 以比例法药敏结果为金标准, 基因芯片法检测利福平耐药的敏感度为 96.1%（73/76）, 特异度为 100.0%（50/50）, 总准确度为 97.6%（123/126）; 检测异烟肼耐药的敏感度为 90.8%（69/76）, 特异度为100.0%（50/50）, 总准确度为 94.4%（119/126）; 基因芯片法检测耐多药结核的敏感度为 86.8%（66/76）, 特异度为100.0%（50/50）, 总准确度为 92.1%（116/126）。 结论 本地区耐多药结核分枝杆菌耐药相关基因优势突变类型为katG 基因 315 位点和rpoB 基因 531 位点, 基因芯片法可快速、有效诊断耐多药结核病。     

Contribution of AGC to ACC and other mutations at codon 315 of the katG gene in isoniazid-resistant Mycobacterium tuberculosis isolates from the Middle East

The presence of ACC and other mutations at codon 315 in the katG gene was detected by PCR amplification followed by restriction fragment length polymorphism (PCR-RFLP) generated with restriction enzymes Msp I and MspA1 I in 37 isoniazid-resistant and 22-susceptible Mycobacterium tuberculosis isolates from Kuwait obtained in 2001. The mutation AGC to ACC was detected in 22 (60%) isolates while any mutation at codon 315 of the katG gene was present in 24 (65%) of 37 isoniazid-resistant isolates. The typing studies showed that majority of the isolates carrying mutations at codon 315 exhibited unique DNA banding patterns. The results were extended by additional analysis of 67, 28 and 17 isoniazid-resistant and 18, seven and six-susceptible M. tuberculosis isolates from Kuwait, Dubai and Beirut, respectively, that were analyzed previously for ACC mutation alone. These studies showed that one of 21, one of 10 and two of 11 isolates (all recovered from patients of Middle Eastern origin) with no AGC to ACC mutation from Kuwait, Dubai and Beirut, respectively, contained other mutations at codon 315 of the katG gene. None of the susceptible strains contained any mutation at codon 315. The PCR-RFLP with MspA1 I that detects all mutations at codon 315, compared with Msp I that detects only ACC mutation, identified more isoniazid-resistant strains with mutations at codon 315 in the katG gene. The data also showed that mutations other than AGC to ACC at codon 315 in the katG gene occur frequently in M. tuberculosis isolates recovered from Middle Eastern patients and should be incorporated in a rapid screen for the detection of mutations for isoniazid-resistance in the katG gene from this ethnic group.     

结核分枝杆菌katG和inhA基因突变的研究

为了解结核分枝杆菌耐异烟肼（ＩＮＨ）分离株ｋａｔＧ基因和ｉｎｈＡ调节序列的突变情况，探讨其在ＩＮＨ耐药性中的可能作用。应用聚合酶链反应（ＰＣＲ）和ＰＣＲ－单链构象多态性（ＳＳＣＰ）方法对６２株结核分枝杆菌临床分离株的ｋａｔＧ基因和ｉｎｈＡ调节序列进行分析。ＰＣＲ分析结果显示：ｋａｔＧ基因和ｉｎｈＡ调节序列的敏感性为１～１０ｐｇＤＮＡ；其引物ＰＣＲ扩增都具有较高的特异性。以Ｈ３７Ｒｖ标准株为对照，１３株敏感株中，１２株ｋａｔＧ基因扩增产物ＳＳＣＰ泳动正常，１株ｋａｔＧ异常；２４株耐非ＩＮＨ抗结核药物株中，８株ｋａｔＧＳＳＣＰ异常；上述２７株的ｉｎｈＡ调节序列ＳＳＣＰ均泳动正常。２５株耐ＩＮＨ株中，３株ｋａｔＧ基因ＰＣＲ扩增阴性，２２株ｋａｔＧ扩增阳性，其中１６株ＳＳＣＰ泳动异常；４株ｉｎｈＡ调节序列ＳＳＣＰ泳动异常，其ｋａｔＧ基因ＳＳＣＰ也异常。结果表明，某些结核分枝杆菌ＩＮＨ耐药性可能是由于其ｋａｔＧ基因完全缺失、ｋａｔＧ基因或ｉｎｈＡ调节序列突变所致，但某些菌株也可能与其无关，是其它耐药基因所致或存在多种机制，尚需进一步探讨。     

结核分枝杆菌异烟肼耐药基因katG点突变的快速检测

目的：探讨快速检测结核分枝杆菌异烟肼（INH）耐药基因型的分子药敏方法。方法：用聚合酶链反应－单链构象多态性（PCR－SSCP）检测了30株INH敏感菌株及30株耐药株的katG基因，随后用SSCP方法鉴定扩增产物有无突变，以结核分枝杆菌H37Rv作对照。结果：所有INH敏感株均观察到katG基因扩增产物，30株INH耐药株中28株观察到katG基因扩增产物。以H37Rv标准株为对照，30株敏感株SSCP带谱与对照相同；28株INH耐药株中13株与对照相同，15株有不同程度的差异。与药敏试验比较，PCR－SSCP检测INH耐药结核菌的敏感性为57%，特异性为100％。结论：多数结核分枝杆菌耐INH是由于katG基因突变所致，用PCR－SSCP筛选突变株可达到快速检测结核分枝杆菌INH耐药基因型的目的。     

Molecular characterization of isoniazid-resistant Mycobacterium tuberculosis clinical strains isolated in the Philippines

The prevalence of mutations in the katG, inhA and oxyR-ahpC genes of isoniazid (INH)-resistant Mycobacterium tuberculosis isolates in the Philippines were determined. Of 306 M. tuberculosis isolates studied, 81 (26.5%) exhibited INH-resistance. Forty-four strains (54.3%) had mutations in the katG gene, eighteen strains (22.2%) had mutations in the putative inhA locus region, seven had mutations in both regions and five strains had mutations in the oxyR-ahpC operon. Only seven strains had no mutations. A total of 71 of the 81 (87.6%) resistant strains and 65 of the 72 (90.3%) INH sensitive randomly selected strains showed amino acid substitution in codon 463 (Arg to Leu) (88.9%). This fact supports the hypothesis that mutations at codon 463 are independent of INH-resistance and are linked to the geographical origins of the strains.     

耐多药结核分枝杆菌三种基因的快速检测

目的 :探讨耐多药结核分枝杆菌耐药基因突变与耐药性的关系。方法 :采用 PCR和 PCR- DS技术对 5 7例耐多药结核临床分离株进行 kat G、rpo B和 emb B基因检测和序列分析。结果 :耐 INH(kat G)、RFP(rpo B)、EMB(emb B)基因突变率分别为 6 3.8%、90 .7%、37.1% ,其中同时耐 INH(kat G)和 RFP(rpo B)基因突变率为 5 4 .1% ,同时耐三种药的基因突变率为 6 5 .6 %。结论 :PCR- DS法对耐两种或两种以上药物的结核检出率较高 ,与传统药敏试验互相弥补 ,对临床用药有指导意义。     

PCR-SSCP技术直接快速检测痰标本中结核分支杆菌rpoB,katG,ahpC基因突变的研究

目的：应用PCR－SSCP技术直接快速检测痰标本中结核分支杆菌rpoB,katG及ahpC基因突变,评价此方法用于结核分杆菌利福平（RFP）及异烟肼（INH）耐药性试验的意义。方法：以结核分支杆菌H37Rv为对照,30例耐RFP（单耐或多耐）、21例耐INH（单耐和多耐）的结核分枝杆菌临床分离株及10例敏感菌株;39例菌阳肺结核患者及30例非结核 性肺部疾病患者痰标本,采用PCR－SS－CP技术检测痰标本和菌株中的结核杆菌rpoB,katG,ahpC基因突变,并与药敏试验对照。结果：PCR－SSCP检测痰标本中结核分支杆菌rpoB,katG及ahpC基因突变的阳性率分别为79％（31／39）、46％（18／39）及5％（2／39）.结论：PCR－SSCP可望成为直接检测临床痰标本中结核分枝杆菌耐利福平及耐异烟肼的快速方法。     

用分子生物学技术快速检测耐多药结核分枝杆菌

目的：用PCR—SSCP技术检测耐INH、RFP、SM结核分枝杆菌分离株KatG、rpoB、rpsL基因突变，评价其在快速检测结核分枝杆菌耐药性方面的临床价值。方法：以结核分枝杆菌H；vRv为对照，20株耐多药菌株和20株敏感株，分别用PCR—SSCP方法检测KatG、rpoB、rpsL基因突变，并将SSCP结果与药敏试验结果作对照分析。结果：PCR—SSCP方法检测20株敏感株SSCP带语均与H37RV标准株相同，而20株耐多药结核分枝杆菌KatG、rpoB、rpsL基因突变的阳性率分别为70％，100％和75％。发现20株耐多药菌株中有11株(55％)共INH、RFP、SM3种耐药遗传标志均发生突变，7株(35％)有2种耐药遗传标志突变，2株(10％)只有RFP耐药标志突变。结论：PCR—SSCP方法敏感、特异，可快速检测结核分枝杆菌KatG、rpoB、rpsL耐药基因突变，有利于耐多药结核分枝杆菌耐药性的快速检测。     

基因芯片技术检测结核分枝杆菌对利福平和异烟肼耐药性的研究

目的探讨基因芯片技术在检测结核分枝杆菌利福平和异烟肼耐药性的应用。方法收集21例临床诊断结核病患者体液标本经培养得到的分离菌液,进行基因芯片检测,包括利福平的rpoB基因位点和异烟肼的KatG和inhA基因位点,从而判断其耐药性。结果 21例样本中,利福平rpoB基因有10例突变,总突变率为47.6%,其中9例为531位点的（C-T）突变,1例526位点的（C-G）突变。与药敏试验的符合率为81.0%。异烟肼基因突变有6例,总突变率为28.6%,其中4例为KatG 315（G-C）突变,2例为inhA-15（C-T）突变。与药敏试验的符合率为90.5%。利福平的11例耐药株中,基因芯片检测8例为突变型,突变率为72.7%,在利福平的10例敏感株中,基因芯片检测2例为突变型,突变率为20.0%。在异烟肼的6例耐药株中,基因芯片检测5例为突变型,突变率为83.3%,在异烟肼的15例敏感株中,基因芯片检测1例为突变型,突变率为6.7%。结论基因芯片技术检测结核分枝杆菌对利福平和异烟肼的耐药性具有较高的特异性和敏感性,快速准确,可以应用于结核分枝杆菌耐药性的检测。     

结核分支杆菌耐药基因检测结果分析

目的 分析肺结核患者结核分支杆菌获得性耐药情况,比较两种耐药性检测方法的检测效果.方法 用药敏试验(绝对浓度法)检测结核分支杆菌株对利福平(RFP)、异烟肼(INH)、链霉素(SM)、吡嗪酰胺(PZA)和乙胺丁醇(EMB)的耐药情况,采用聚合酶链反应-单链构象多态性分析(PCR-SSCP)检测结核分支杆菌耐药rpoB、katG、rpsL、pncA和embB基因突变.结果:400例肺结核耐药患者常规药敏试验耐药316例(79.8%);耐药基因检测耐药株288株,突变率72.0%.RFP耐药例数和耐药率明显高于SM、PZA和EMB(耐药例数:F=2.45,2.56,2.69,P＜0.05;耐药率:F=2.55,2.66,2.79,P＜0.05),rpoB突变株和突变率明显高于katG、rpsL、pncA和embB(突变株:F=2.28,2.46,3.19,3.33,P＜0.05～0.01;突变率:F=2.36,2.61,3.25,3.45,P＜0.05～0.01),高浓度药物耐药菌株的突变株和突变率显著高于低浓度药物耐药菌株,随着不规律用药时间的延长,耐药率和突变率均逐渐上升(耐药率:F=2.77,2.88,P＜0.05;突变率:F=2.72,2.85,P＜0.05).结论 PCR-SSCP是一种快速检测结核杆菌rpoB、katG、rpsL、pncA和embB基因突变敏感、特异的方法.     

广东地区结核分枝杆菌利福平耐药与基因rpoB突变关系分析

目的 为了解广东地区结核分枝杆菌rpoB基因突变特点,并探讨其与利福平耐药之间的关系.方法 本研究根据结核病诊断细菌学检验规程从临床结核病疑似患者痰液中分离鉴定结核分枝杆菌,采用绝对浓度法对其进行药敏试验,并用PCR-DNA直接测序法对结核分枝杆菌rpoB基因进行序列分析.结果 结果分离得到利福平耐药株27株,其中低浓度耐药8株,高浓度耐药19株.经测序分析,所有利福平敏感株rpoB基因均未发生氨基酸突变 利福平耐药株rpoB基因的突变率为81.5%（22/27）,突变均发生在利福平耐药决定区内,突变位点为526、531及533,以Ser531Leu突变最为常见（55.6%,15/27）,并且高浓度耐药株的突变率明显高于低浓度耐药株.结论 结果表明广东省结核分枝杆菌rpoB基因突变是结核分枝杆菌利福平耐药性产生的主要分子机制,突变位点同国内外研究基本一致,但各位点突变形式所占比例有自身特点.     

Mechanisms of isoniazid resistance in Mycobacterium tuberculosis.

Isoniazid (INH) is a widely used front-line antituberculous agent with bacteriocidal activity at concentrations as low as 150 nM against Mycobacterium tuberculosis. INH is a prodrug and requires activation by an endogenous mycobacterial enzyme, the catalase-peroxidase KatG, before exerting toxic effects on cellular targets. Resistance to INH develops primarily through failure to activate the prodrug due to point mutations in the katG gene. In addition to mutations in katG, mutations in several other loci, such as the alkylhydroperoxidase AhpC and the enoylreductase InhA, may contribute to INH resistance. Although these markers can be used to accurately predict clinical INH resistance in a large number of cases, the molecular mechanisms involved remain largely speculative and incomplete.     

耐氟喹诺酮类铜绿假单胞菌gyrA及parC基因突变研究

目的　研究临床分离的耐氟喹诺酮类铜绿假单胞菌gyrA及parC基因突变情况。方法　测定临床分离的 5 5株铜绿假单胞菌MIC值 ,从中筛选出 1株敏感菌和 8株耐药菌 ,以标准敏感菌株ATCC2 785 3作为质控菌株。用聚合酶链反应 (PCR)扩增gyrA及parC基因的喹诺酮耐药决定区 (QR DR) ,扩增产物片段长度分别为 35 1bp、397bp。用限制性内切酶SacⅡ消化gyrAPCR产物 ,同时对上述 10株菌的gyrA及parC基因的喹诺酮决定区 (QRDR)进行PCR DNA直接测序分析。结果　有 8株耐菌株的gyrA基因在 83位 (ACC→ATC)有突变 ,导致氨基酸Thr→Ile的改变 ;有 3株高度耐药菌gyrA基因同时在 87位 (GAC→GGC)有突变 ,导致氨基酸Asp→Gly的改变 ;有 4株耐药菌株的parC基因在 87位有TCG→TTG突变 ,导致氨基酸由Ser→Leu的改变。同时具gy rA和parC突变MIC值是仅具gyrA突变菌株MIC值的 2～ 16倍。未发现parC突变单独存在。另外 ,有 6株耐药菌gyrA的 132位有CAC→CAT的突变 ;所有耐药菌株parC基因 115位有GCT→GCG的突变 ,该突变未引起氨基酸的改变。结论　gyrA83、87位突变及parC基因 87位突变都可引起铜绿假单胞菌对氟喹诺酮类药物产生耐药 ,但以gyrA基因 83位突变为主 ,合并gyrA基因 87位及parC基因 87位突变可增加耐药程度。     

Detection of novel coupled mutations in the katG gene (His276Met, Gln295His and Ser315Thr) in a multidrug-resistant Mycobacterium tuberculosis strain from Chennai, India, and insight into the molecular mechanism of isoniazid resistance using structural bioinformatics approaches

This study reports on the structural basis of drug resistance targeting the katG gene in a multidrug-resistant Mycobacterium tuberculosis (MDR-TB) strain with two novel mutations (His276Met and Gln295His) in addition to the most commonly reported mutation (Ser315Thr). A structural bioinformatics approach was used to predict the structure of the mutant KatG enzyme (MT). Subsequent molecular dynamics and docking studies were performed to explain the mechanism of isoniazid (INH) resistance. The results show significant conformational changes in the structure of MT leading to a change in INH binding residues at the active site, with a significant increase in the inhibition constant (Ki) of 5.67 μm in the mutant KatG-isoniazid complex (MT-INH) compared with the wild-type KatG-isoniazid complex (WT-INH). In the case of molecular dynamics studies, root mean square deviation (RMSD) analysis of the protein backbone in simulated biological conditions revealed an unstable trajectory with higher deviations in MT throughout the simulation process (1 ns). Moreover, root mean square fluctuation (RMSF) analysis revealed an overall increase in residual fluctuations in MT compared with the wild-type KatG enzyme (WT), whilst the INH binding residues of MT showed a decreased fluctuation that can be observed as peak deviations. Hence, the present study suggests that His276Met, Gln295His and Ser315Thr mutations targeting the katG gene result in decreased stability and flexibility of the protein at INH binding residues leading to impaired enzyme function.     

铜绿假单胞菌临床分离株gyrA基因突变与喹诺酮类药物耐药关系研究

目的:研究铜绿假单胞菌临床分离株gyrA基因突变与喹诺酮类药物耐药关系,并对聚合酶联反应(PCR)-限制性片段长度多态性(RFLP)-DNA单链构象多态性(SSCP)分析铜绿假单胞菌临床分离株gyrA基因突变的可行性进行评估。方法:以铜绿假单胞菌临床分离株gyrA基因序列为靶序列,用PCR、PCR-RFLP、PCR-SSCP、DNA测序等方法对铜绿假单胞菌ATCC27853及16株临床分离株gyrA基因突变进行对比研究。结果:在8株耐环丙沙星铜绿假单胞菌中,有6株gyrA基因的83位表现出单点突变,其突变方式全为ACC→ATC,导致氨基酸苏氨酸→异亮氨酸的改变;gyrA基因的PCR扩增产物SacⅡ酶切片段与测序结果一致;SSCP分析结果显示,16株细菌中仅2株gyrA带型与ATCC27853相同,其它菌株gyrA带型与ATCC27853均不同。结论:临床分离的铜绿假单胞菌对喹诺酮类药物耐药的分子机制主要表现为gyrA基因83位氨基酸密码子突变,应用PCR-RFLP-SSCP系统可快速、准确地检测耐喹诺酮类药物的铜绿假单胞菌gyrA中碱基的变异。     

Comparison of gyrA gene mutations between laboratory-selected ofloxacin-resistant Mycobacterium tuberculosis strains and clinical isolates

To understand the relationship between mutations in the quinolone resistance-determining region (QRDR) of the gyrA gene and drug resistance to ofloxacin, 85 laboratory-selected ofloxacin-resistant Mycobacterium tuberculosis mutant strains and 110 M. tuberculosis clinical isolates, screened by denaturing high-performance liquid chromatography to contain mutations, were analysed for their mutation patterns by sequencing as well as their ofloxacin minimal inhibitory concentrations (MICs). All mutations detected occurred at the codons Ala74, Ala90, Ser91 and Asp94 in all strains. One of the five different forms of missense mutation in Asp94 occurred in 60% of the laboratory-selected strains and 78% of the clinical isolates. However, 53 clinical isolates (48%) and only 2 laboratory-selected strains (2.4%) harboured double point mutations. The mutation Ala74Ser occurred only in the clinical isolates and only in combination with the Asp94Gly mutation. The ofloxacin MIC for the clinical isolates ranged from 0.5microg/mL to 20microg/mL, whilst the MICs for the laboratory-selected strains were > or =10microg/mL. The differences in gyrA gene mutation patterns and MICs between the laboratory-selected resistant strains and clinically isolated resistant strains identified here might help to understand the mechanisms involved in fluoroquinolone resistance.     

The occurrence of rare rpoB mutations in rifampicin-resistant clinical Mycobacterium tuberculosis isolates from Kuwait

Mutations associated with rifampicin (RIF) resistance in two regions of the rpoB gene were studied by line probe (INNO-LiPA Rif. TB) assay (LiPA) and/or DNA sequencing in 32 RIF-resistant and 21 RIF-susceptible Mycobacterium tuberculosis strains isolated from 53 tuberculosis patients in Kuwait. The LiPA identified all susceptible strains as RIF sensitive, and the DNA sequences of the 81bp rifampicin resistance-determining region (RRDR) and the N-terminal region of the rpoB gene from five isolates were identical to the wild-type sequences from M. tuberculosis H(37)Rv. The LiPA identified 24 of 32 (75%) phenotypically documented RIF-resistant M. tuberculosis isolates as RIF resistant, with specific detection of mutation in 19 isolates, whilst 8 strains were identified as RIF susceptible. DNA sequencing of the RRDR confirmed the results for the 19 RIF-resistant isolates and identified precise mutations in five isolates for which specific base changes could not be determined by LiPA, as well as identifying a mutation (insertion 514TTC) in three isolates and no mutation in five of the eight isolates that were detected as RIF susceptible by LiPA. Two of the latter five isolates contained a V146F mutation in the N-terminal region of the rpoB gene and were recovered from patients of Middle Eastern origin. These analyses identified 11 different mutations in two regions of the rpoB gene. The majority of the isolates carrying identical or no mutations in the rpoB gene exhibited unique DNA banding patterns in fingerprinting studies. The occurrence of rare mutations in the rpoB gene, namely insertion 514TTC in 3 (9%) and V146F in 2 (6%) of 32 RIF-resistant M. tuberculosis isolates from Kuwait, is the highest reported so far.     

Structure‐Based Design of a Novel Class of Potent Inhibitors of InhA,the Enoyl Acyl Carrier Protein Reductase from Mycobacterium Tuberculosis: A Computer Modelling Approach

The NADH‐dependent Enoyl‐ACP reductase (InhA) of Mycobacterium tuberculosis has been shown to be the primary target of the frontline drug isoniazid (INH). However, INH must be first activated by katG gene, mutations in which have mediated resistance to INH. Recently, direct inhibitors of InhA have been reported. Using a structure‐based approach, we have identified a tripeptide inhibitor with the sequence WYW, which is 100 times more potent than the existing inhibitors. It is therefore, a potential lead compound for the development of new anti‐TB drugs.     

耐药基因检测在初次复治肺结核患者中的应用价值

目的 研究耐药基因检测在初次复治肺结核中的临床价值.方法 选取河北省胸科医院2014年1月-2015年12月收治的初次复治时痰液或支气管肺泡灌洗液浓缩集菌抗酸染色阳性的肺结核患者128例,采用BACTEC MGIT 960培养基对痰标本进行培养,同时分别选取高浓度和低浓度异烟肼(INH)和利福平(RFP)进行药敏试验,并采用基因芯片法对药敏阳性菌株进行耐药基因突变检测,分析药敏阳性菌株的耐药基因突变率.结果 128例初次复治肺结核患者中,对RFP的耐药率为25.00％,对INH的耐药率为22.66％.基因芯片检测耐RFP结核分枝杆菌的rpoB基因的符合率为96.23％、特异性为97.46％、敏感度为91.67％;基因芯片检测耐INH结核分枝杆菌的katG基因的符合率为94.89％、特异性为98.14％、敏感度为81.94％.结论 对初次复治肺结核患者耐药基因的检测,可快速、准确的检测出结核分枝杆菌耐药基因,准确推断耐药种类,为临床早期诊断及治疗提供科学指导.