PCR-SSCP快速检测耐多药结核分枝杆菌

目的:了解本地区结核病耐药基因突变情况,探讨PCR-SSCP作为新的分子药敏试验方法在临床的应用价值。方法:通过提取耐INH、RFP、SM的肺结核患者痰中结核分枝杆菌DNA,进行PCR-SSCP分析,检测结核分枝杆菌rpoB、katG、rpsL基因是否存在突变,并与传统L-J药敏实验对照。结果:30株耐多药株中,耐RPF、INH、SM基因突变阳性率为90%(27/30)、63%(19/30)、53%(16/30)。3个基因联合突变共8株(26.7%),2个基因联合突变共18株(60%),即26株(86.7%)。单基因突变共2株,2株无基因突变。结论:通过PCR-SSCP方法可检测出绝大部分耐多药结核病的耐药基因,rpoB、katG、rpsL基因突变与本地区结核杆菌对RFP、INH、SM耐药性有关。与传统L-J药敏实验对比,PCR-SSCP是一种敏感、快速的指导临床用药的先进检测方法。     

Detecting katG Drug Resistant Genetic Mutation among pneumoconiosis patients complicated with tuberculosis in Mycobacterium tuberculosis L-forms by PCR-SSCP

Objective：To study the relationship between mutation in the katG gene and drug resistance of INH in Mycobacterium tuberculosis L-forms among patients of pneumoconiosis complicated with pulmonary tuberculosis, and to explore the clinical application of PCR-SSCP. Methods： A total of 52 clinical isolated strains of Mycobacterium tuberculosis L-forms were collected. Mutation in the katG genes was detected by PCR-SSCP and traditional antimicrobial susceptibility test （AST）. Results： The results by AST showed that there were 40 persisters in 52 clinical isolated strains. The drug resistant rate was 76.92%（40/52）, and the gene mutation rate of katG was 57.70%（30/52）by PCR-SSCP, the difference was no quite significance （X^2 = 2.8507, P 〉 0.05）. The coincidence rate of two methods was 75.00% （30/40）. Conclusion： The detectionrate of katG resistant strains in Mycobacterium tuberculosis L-forms was high by PCR-SSCP. The combined application of PCR-SSCP and traditional antimicrobial susceptibility test can improve the detecting rate.     

Application of PCR-SSCP in detecting rpoB drug resistant gene polymorphism of M. tuberculosis L-form from pneumoconiosis patients with tuberculosis

Objective: To study the relationship between the polymorphism of drug resistant gene rpoB and drug resistance against rifampicin(RFP) of M. tuberculosis L-forms, and to evaluate its clinical application. Methods: A total of 52 clinical isolated strains of M. tuberculosis L-forms were collected. rpoB gene polymorphism was analyzed by polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) and conventional antimicrobial susceptibility test (AST). Their results were compared. Results: AST results showed that 38 of 52 clinical isolated strains were drug resistance (73.08％),while PCR-SSCP indicated 65.38％ (32/52) rpoB gene polymorphism. There was no statistic significance(χ2= 2.4914) between the 2 methods. Conclusion:Combined the application of PCR-SSCP with AST in detecting rpoB drug resistant gene polymorphism of M. tuberculosis L-form from pneumoconiosis patients with tuberculosis may have advantages at earlier diagnosis and guidance of clinical medications.     

Molecular mechanisms of drug resistance in Mycobacterium tuberculosis clinical isolates

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耐多药结核分枝杆菌基因突变在耐药性检测中的应用

目的:探讨结核分枝杆菌耐链霉素(SM)和利福平(RFP)的耐药分子机制,应用聚合酶链反应一单链构象多态性(PCR-SSCP)同时检测rpsL基因、rpoB基因突变在结核分枝杆菌耐SM和RFP耐药性测定中的应用价值。方法:采用PCR-SSCP技术对168株结核分枝杆菌临床分离株rpsL基因、rpoB基因突变同时进行检测。即首先用PCR方法同时扩增RFP、SM的rpoB基因和rpsL基因,然后进行SSCP分析。结果:以结核分枝杆菌H37RV为对照,82株药物敏感株的rpsL基因、rpoB基因均未见SSCP图谱异常。特异性为100%。86株同时耐SM和RFP的耐药株中,68株(79.1%)有rpsL基因图谱异常;81株(94.2%)有rpoB基因图谱异常。结论:rpoB基因突变和rpsL基因突变分别是结核分枝杆菌耐RFP和SM的主要分子机制。应用PCR-SSCP技术可同时快速检测结核分枝杆菌SM和RFP耐药性。     

PCR-SSCP检测耐多药结核菌基因与细菌培养结果对比分析

目的应用PCR-SSCP检测常规及BACTEC培养耐INH、RFP、SM的耐多药结核分枝杆菌临床分离株KatG、rpoB、rpsL基因突变,分析其敏感性和特异性,评价其在检测结核分枝杆菌耐药性方面的价值和临床实用性。方法22株常规培养和8株BACTEC培养检出的高浓度及低浓度耐INH、RFP、SM耐多药结核分枝杆菌分离株,分别用PCR-SSCP检测KatG、rpoB、rpsL基因,观察其电泳条带,并与结核菌标准株H37RV对比。结果22株常规培养耐INH、RFP、SM的耐多药结核分枝杆菌分离株中,KatG、rpoB、rpsLPCR扩增产物,用SSCP分别检出KatG基因16株(72.7%)、rpoB基因19株(86.4%)和rpsL基因14株(63.6%)。8株BACTEC培养耐INH、RFP、SM的耐多药结核分枝杆菌分离株中分别检出KatG基因5株(62.5%),rpoB基因6株(75%)、rpsL基因5株(62.5%)。综合两者KatG、rpoB和rpsL基因检出率分别为70.0%(21/30)、83.3%(25/30)和63.3%(19/30)。高浓度耐药与低浓度耐药分离株中的检出率有显著差异(P<0.05),常规培养与BACTEC培养分离株中的检出率无显著差异(P>0.05),与结核菌标准株H37RV电泳条带对比,特异性为100%,敏感性97%。结论PCR-SSCP敏感、特异,可快速检测结核分枝杆菌的KatG、rpoB和rpsL基因,可用于耐多药结核分枝杆菌的临床检测。     

PCR-SSCP法检测结核分枝杆菌耐药性

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以及聚合酶链反应-单链构象多态性分析(poly merase chain reaction-single strand cinfomlation polymorphism，PCR-SSCP)方法的临床应用价值。方法：用PCR-SSCP方法检测58株结核分枝杆菌临床分离株katG，rpoB，rpsL基因突变并与常规药敏试验检测结果进行对比。结果：经常规药敏试验检测，58株结核分枝杆菌临床分离株中共有41株耐药，其中，耐异烟肼(INH)为35株，高耐药株27株；耐利福平(RFP)为31株，高耐药株24株；耐链霉素(SM)有31株，高耐药株26株。同时耐3种药物的有21株(51．2％)，耐2种药物的14株(34．1％)，单耐药株6株(14．6％)．PCR-SSCP方法对58株临床分离株katG，rpoB，rpsL基因突变的检测率为40％(23／58)，45％(26／58)，38％(22／58)，其中检出3个基因同时突变的有13株(32％)，2种基因突变的12株(29％)，1种基因突变的有10株(23．8％)．常规药敏试验与PCR-SSCP法检出结核分枝杆菌同时耐3种药物的符合率为61．9％(13／21)，检出耐2种药物的符合率为85．70k，(12／14)，检出耐1种药物的符合率为50％(3／6)．高耐药株中突变率为80．5％(62／77)，低耐药株中突变率为60％(12／20)．结论：PCR-SS-CP方法对耐2种以上药物的结核杆菌检出率较高，且耐药基因突变率随着耐药浓度增高而增高。将PCR-SSCP法与药敏试验联合应用可互相弥补，已成为临床指导用药的好方法。     

PCR-DNA序列测定法检测脊柱结核分枝杆菌耐药基因的研究

目的 研究聚合酶链反应、DNA序列测定在脊柱结核分枝杆菌耐药性临床应用的价值.方法 对25例脊柱结核病例脓液、干酪样坏死组织,采用PCR法扩增样本的耐药基因rpoB、rpsL、katG片段 以PCR-DNA序列测定法检测扩增产物,利用DNATools（5.1）软件、NCBI/BLAST公共数据库对耐药基因比较分析.结果 73份耐药基因片段, 检测出15份基因突变,其中rpsL基因突变4份,rpoB基因突变5份,katG基因突变6份,平均报告时间6d.结论 脊柱结核分枝杆菌耐药基因序列检测具有快速、灵敏和特异性高等特点,与细菌学等方法相结合,能为脊柱结核临床治疗提供有利的帮助.     

Molecular characteristics of rifampin and isoniazid resistant Mycobacterium tuberculosis strains from Beijing, China

Background China is one of the high burden countries of Mycobacterium tuberculosis （TB） infection globally, with high incidence and mortality. We studied the molecular characteristics of rifampin （RIF） and isoniazid （INH） resistant Mycobacterium tuberculosis strains from Beijing, China, in order to find out the genetic marker for rapid detection of specific drug resistance. Methods Forty pansusceptible and 81 resistant strains of Mycobacterium tuberculosis isolated from Beijing, China during 2002-2005 were analyzed. The modified rifampin oligonucleotide （RIFO） assay based on reverse line blot hybridization was used to detect mutations in the 81 bp hot-spot region of rpoB gene, which is associated with RIF resistance. The INH resistance associated genes, regulatory region mab-inhA （-15C/T） and structural gene katG S315T were detected by reverse line blot hybridization and PCR-restriction fragment length polymorphism （RFLP） method respectively. All the strains were typed by spoligotying and the Beijing genotype was further subdivided by NTF locus analysis. The distribution of drug resistance associated mutations in the above genes was compared in these groups. Results Sixty-five （91.5%） of 71 RIF resistant and 52 （92.9%） of 56 multidrug-resistant （MDR, i.e. resistant to at least RIF and INH） strains were found to harbor mutations in the rpoB hot-spot region. No mutation was detected in RIF sensitive strains. The specificity and sensitivity of the modified RIFO assay were 100% and 91.5%, respectively, katG315 AGC〉ACC and inhA-15C〉T mutations were found in 40 （60.6%） and 10 （15.2%） of 66 INH resistant strains, respectively; 7.6% of INH-resistant strains had mutations in both of these genes. Therefore, a combined use of both katG315 and inhA-15 identified 68.2% of INH-resistant strains. The Beijing genotype accounted for 91.7% of total strains and was further subdivided into ＂modern＂ （76.6%） and ＂ancestral＂ （23.4%） group. There is no significant difference between ＂ancestral＂ and ＂modern＂ group in prevalance of drug resistance-associated gene mutations. Conclusions The hot-spot region of rpoB gene can be used as genetic marker for detection of RIF resistant strains; a combined use of both katG315 and inhA-15 can improve the detection rate of I NH resistant strains; the Beijing genotype is prevalent in Beijing, China; the modified RIFO assay can be a practical tool for rapid detection of RIF resistant and MDR isolates in the routine diagnostic work.     

深圳地区结核临床分离菌基因突变与耐药的关系

目的研究深圳地区结核分枝杆菌临床分离株的基因突变与利福平（RFP）、异烟肼（INH）、链霉素（SM）和乙胺丁醇（EMB）耐药之间的关系。方法采用反向斑点杂交技术（RDBHA）对182株结核分枝杆菌临床分离株的rpoB、katG、inhA、rpsL和embB基因突变位点进行检测,并采用L-J比例法检测这些分离株对RFP、INH、SM、EMB的耐药性。结果 182株临床分离株中,总的基因突变率为34.62%（63/182）,其中rpoB基因突变率最高,达24.17%（44/182）。多重耐药结核菌（MDR-TB）占17.03%（31/182）。以L-J比例法作为金标准,采用RDBHA技术检测分别与RFP、INH、SM、EMB耐药相关的rpoB、katG/inhA、rpsL、embB基因突变,其灵敏度分别为88.89%、67.50%、81.82%和64.29%,特异度分别为97.08%、94.37%、97.99%和92.86%。结论深圳地区结核分离株ropB基因突变最普遍,S531L位点是其突变率最高的位点。深圳地区结核分枝杆菌对4个一线抗痨药物耐药现象均较严重,反向斑点杂交技术（RDBHA）可快速检测结核分枝杆菌耐药基因突变,能给临床提供快速用药指导。     

临床肺结核治疗中耐药性分析及对策

目的:了解从分离自肺结核患者的结核分枝杆菌耐药基因突变率及耐药情况,以利抗结核治疗中药物的合理选用。方法:对痰涂片阳性的新发初治和复治肺结核患者进行痰结核分枝杆菌培养,阳性菌株采用高、低两种药物浓度,四种抗结核药物耐药性测试,同时用实时PCR法对结核分枝杆菌的耐药基因和突变进行检测。结果:127例痰培养阳性菌株总耐药率为14.2%,其中初治耐药率8.1%,复治耐药率35.7%,对四种抗结核药物的耐药率依次为异烟肼8.7%,利福平2.4%,链霉素2.4%,乙胺丁醇0.8%。耐药基因检测结果,在初治组中rpoB和katG的突变率为12.1%(12/99)和10.1%(10/99);复治组中rpoB和katG的突变率为32.1%(9/28)和21.4%(6/28)。结论:分离自肺结核患者的结核分枝杆菌在初始治疗时已存在耐药性,而药物治疗有可能使其耐药性增加。结果表明抗结核治疗前及在治疗过程中对结核分枝杆菌进行耐药性及耐药基因检测对指导临床抗结核治疗很有实际意义。     

结核分支杆菌rpoB基因突变和耐利福平的相关性研究

研究结核分支杆菌ｒｐｏＢ基因突变及其与利福平（ＲＦＰ）耐药性的关系,建立快速检测耐药基因型的分子药敏试验方法。用ＰＣＲ—ＳＳＣＰ分析结核分支杆菌ｒｐｏＢ基因。结果显示ＰＣＲ扩增ｒｐｏＢ基因为属特异性。８１株结核分支杆菌的ＰＣＲ－ＳＳＣＰ图谱与Ｈ３７Ｒｖ为对照,３５株敏感菌仅有一株位置有区别;４６株耐药菌有４２株有明显区别;检测阳性率为９１．３％;特异性９７．１％。证明ｒｐｏＢ基因突变是结核分支杆菌耐利福平的主要机理,ＰＣＲ—ＳＳＣＰ可简便快速地检出ｒｐｏＢ基因的突变,有助于结核分支杆菌耐药性的快速检测和研究。     

浙东地区耐多药结核分枝杆菌耐药特性及分子机制研究

目的 对浙东地区耐多药结核分枝杆菌（MDR-TB）耐药性及分子机制进行研究,为治疗耐多药结核病提供理论依据。方法 收集2018 年1 月～2019 年12 月浙江东部9 家结核病定点医院临床分离的结核分枝杆菌。采用比例法检测异烟肼（INH）、利福平（RIF）、氧氟沙星（OFL）、链霉素（SM）、乙胺丁醇（EMB）、阿米卡星（AK）、对氨基水杨酸（PAS）、卷曲霉素（CM）和丙硫异烟胺（TH）对MDR-TB 的耐药性。通过基因芯片方法检测耐多药结核分枝杆菌rpoB、katG、inhA 突变位点,PCR 扩增OFL 耐药的MDR-TB 的gyr 耐药基因并测序。结果 耐多药结核分枝杆菌对OFL、SM、EMB、AK、PSA、CM、TH、INH 和RIF 耐药率分别为38.1%、54.8%、28.6%、11.9%、8.3%、9.5%、13.1%、100.0%和100.0%。耐多药结核分枝杆菌的突变位点为rpoB 511（9 例）、rpoB 513（3 例）、rpoB 516（3 例）、rpoB 526（25 例）、rpoB 531（38 例）、rpoB 533（2 例）,KatG 315（71 例）,inhA-15(4 例),KatG 315 与inhA-15 同时突变（9 例）。检测到26 株gyrA 基因和2 株gyrB 基因发生突变,突变类型为Thr478Asn、Asn477Thr、Ala90Val、Ser91Pro、Asp94Ala、Asp94His、Asp94Asn 和Asp94Gly。结论 耐多药结核分枝杆菌利福平耐药以rpoB 基因531位点突变为主;异烟肼耐药以KatG 基因315 位点突变为主;MDR-TB 对喹诺酮类药物的耐药机制以gyrA 基因Ala90Val、Ser91Pro、Asp94Gly 突变类型为主。     

Molecular Characterization of Drug-Resistant Beijing Family Isolates of Mycobacterium Tuberculosis from Tianjin,China

Objective Tuberculosis remains a severe public health issue, and the Beijing family of mycobacterium tuberculosis （M. tuberculosis） is widespread in East Asia, especially in some areas in China, like Beijing and Tianjin. This study aimed at determining the mutation patterns of drug-resistant Beijing strains of M. tuberculosis isolated from Tianjin, China. Methods A total of 822 M. tuberculosis isolates were screened for drug resistance by an absolute concentration method and the genotype was identified by PCR. 169 drug-resistant isolates of the Beijing family were analyzed for the potential mutations in the rpoB, katG, inhA promoter region and in rpsL, rrs and embB genes, which are associated with resistance to rifampin （RFP）, isoniazid （INH）, streptomycin （SM） and ethambutol （EMB） respectively by PCR and DNA sequencing. Results Fifty-eight out of 63 RFP-resistant isolates were found to carry the mutations within the 81-bp RFP resistance determining region （RRDR） of the rpoB gene and the most frequent mutations occurred at codon 531 （44.4%）, 526 （28.6%）, and 516 （7.9%） respectively. 16 mutation pattems affecting 12 different codons around the RRDR of rpoB were found. Of 116 INH-resistant isolates, 56 （48.3%） had the mutation of katG 315 （AGC→ACC） （Ser→Thr）, 3 （2.6%） carried S315N （AGC→AAC） and 27 （16.0%） had the mutation of inhA-15A→T. 84 out of 122 SM-resistant isolates （68.9%） displayed mutations at the codons 43 or 88 with AAG→AGG （Lys→Arg） of the rpsL gene and 22 （18.0%） with the mutations at positions 513A→C, 516C→T or 905 A→G in the rrs gene. Of 34 EMB-resistant isolates, 6 had mutation with M306V （ATG→GTG）, 3 with M306I （ATG→ATT）, 1 with M306I （ATG→ATA）, 1 with D328Y （GAT→TAT）, 1 with V348L （GTC→CTC）, and 1 with G406S （GGC→AGC） in the embB gene. Conelusion These novel findings extended our understanding of resistance-related mutations in the Beijing strains of M. tuberculosis and may provide a scientific basis for development of new strategies for diagnosis and control of tuberculosis in China and other countries where Beijing strains are prevalent.     

Sequence analysis on drug-resistant rpoB gene of Mycobacterium tuberculosis L-form of isolated from pneumoconlosls workers

To study the characteristics of drug-resistant genetic mutation of rpoB on coal workers＇ pneumoconiosis complicated with L-form of Mycobacterium tuberculosis. Methods： A total of 42 clinical isolated strains of Mycobacterium tuberculosis L-forms were collected, including 31 drug-resistant strains. Their genomes DNA were extracted, the target genes were amplified by PCR, and the hot regions in the rpoB gene were analyzed by automated DNA sequenator. Results： No mutation of rpoB gene was identified in 11 rifampicin-sensitive strains while conformation changes were found in 31 rifampicin-resistant strains. The mutation rate was 93.55% （29/31） in resistant strains, mainly concentrated in codon 531 （51.6%, 16/31） and 526 （32.26%, 10/31）. Base substitutions happened, including 27 unit point mutation and 2 two point mutation. The mutation of codon 516 that new found wasn＇t reported by internal and overseas scholars. Conclusion： The substitution of highly conserved amino acids encoded by rpoB gene results in the molecular mechanism responsible for rifampicin resistance in Mycobacterium tuberculosis L-forms. It also proves that rpoB gene is diversiform.     

耐异烟肼结核分枝杆菌及其katG、inhA基因的突变

目的 探讨耐异烟肼结核分枝杆菌及其katG与inhA基因突变的特征,为临床选择抗结核药物治疗提供实验室参考。方法 回顾性分析2015年1月1日-2017年1月1日在上海交通大学附属同仁医院海南分院的260例患者标本,对213例进行结核分枝杆菌分离培养,对47例进行博奥芯片法检测,再对213例培养所得菌株进行博奥芯片检测,对博奥芯片检测的47例痰标本进行结核分枝杆菌分离培养;对101株结核分杆菌进行katG与inhA基因检测。结果 培养法和博奥芯片结核分枝杆菌检出率分别为36.6%（78/213）和48.9%（23/47）;培养法和博奥芯片耐多药结核分枝杆菌检出率分别为41.0%（32/78）和47.8%（11/23）;博奥芯片法和比例法耐异烟肼符合率98.7%(77/78);男性结核分枝杆菌阳性率45.9%高于女性阳性率29.2%;检测katG基因和inhA基因,3个基因区域都发生突变,突变发生率大小依次为katG315（AGC→ACC）密码子(32株,54.2%)、katG315（AGC→AAC）密码子(16株,27.1%)、inhA-15（C→T）(10株,16.9%)。24株(23.8%,24/101)对异烟肼无突变但对利福平发生突变;43株也同时耐利福平(42.6%,43/101)。突变频率较高的突变位点是315,突变频率为81.4%(48/59),1株(1.7%,1/59)为双位点联合突变且突变频率最低。结论 结核分枝杆菌katG和inhA基因突变与耐INH相关,这为临床及时准确诊断、及早使用抗结核药物联合治疗提供了帮助。     

耐多药结核分支杆菌突变基因检测

目的：研究耐多药结核分支杆菌耐药分子机制,建立快速检测耐药基因型的分子药敏试验方法,方法：通过16S rRNA聚合酶链反应单链的构象多态性（polymerase chain reaction-single strand conformation polymorphism,PCR-SSCP)方法对30株耐多药分离株和20株药物敏感株进行初步分子菌种鉴定。通过PCR－SSCP分析30株耐多药结核病临床分离株的rpoB,katG,rpsL基因。结果：经16S rRNA PCR-SSCP菌种鉴定,所分析菌株均为结核分支杆菌;30株耐多药分离株经SSCP分析,56．7％存在rpoB基因突变,43．3％有katG基因突变,91．3％有rpsL基因突变,结论：rpoB,rpsL,katG基因突变分别是结核分支杆菌耐利福平,链霉素,异烟肼的分子机制,耐多药结核病是药物靶基因突变所致。通过PCR－SSCP可简便,快速地检测大部分耐多药结核病的耐药基因。     

结核分支杆菌耐利福平耐药基因检测研究

目的评价应用DNA序列分析方法和聚合酶链反应-单链构象多态性(PCR-SSCP)技术检测rPOB基因突变在结核分支杆菌耐利福平(RFP)耐药性测定中的应用价值.方法采用DNA序列分析法和PCR-SSCP法对80株结核分支杆菌临床分离株(其中药物敏感株32株,耐RFP或含耐RFP耐多药株48株)rpoB基因核心区域的突变情况进行检测.结果以结核分支杆菌H37RV为对照,所有32株药物敏感株的rPOB基因均无突变.用DNA序列分析方法检测48株耐药株中44株发生突变,敏感性为91.7%(44/48),其中最常见的突变位点是531位丝氨酸和526位组氨酸.PCR-SSCP检测48株耐RFP分离株中,45株rPOB基因SSCP图谱异常,其敏感性为93.1%(45/48).结论DNA序列分析可指导临床用药,PCR-SSCP可快速检测结核分支杆菌RFP耐药性.     

结核分支杆菌rpoB基因套式PCR在利福平耐药和结核病辅助诊断中的应用

目的 分析耐利福平的结核分支杆菌(Mtb)rpoB基因突变状况。方法 对38株耐利福平Mtb菌株的rpoB基因突变高频区域进行套式PCR扩增，并进一步全自动测序。结果 38株耐利福平Mtb中有36株存在突变(包含5个密码子11种形式的突变)，突变率94．7％。结论 Mtb xpoB基因的突变与利福平耐药高度关联，rpoB基因套式PCR及测序能快速、正确鉴别耐利福平菌株。     

结核分支杆菌耐药分子机制的检测技术的研究

目的：探讨结核分支杆菌对异烟肼、利福平耐药的分子机制，建立快速检测耐药分支杆菌基因型的分子生物学方法。方法：采用聚合酶链反应一单链构象多态性(PCR—SSCP)同时检测结核分支杆菌敏感株和耐异烟肼、利福平耐药株的KayG基因、rpoB基因突变。结果：78株结核分支杆菌临床分离株均未发现KatG、rpoB基因缺失。以结核分支杆菌H37RV为对照，36株药物敏感株的KarG、rpoB基因SSCP图谱均正常。42株同时耐异烟肼和利福平的多耐药株中，KatG和rpoB基因突变率分别为66．7％(28／42)，90．5％(38／42)。结论：结核分支杆菌耐异烟肼、利福平耐药性的产生是由于KatG基因和rpoB基因突变所致。应用PCR-SSCP技术可同时快速检测结核分支杆菌异烟肼、利福平耐药性。