空洞型肺结核组织样本耐药菌及相关基因检测的临床研究  

Clinical research on the drug resistance bacteria and related genetic testing in the tissue samples of cavitary pulmonary tuberculosis

作　　者：薛宗锡[1] 刘志辉[2] 谭守勇[3] 尹小毛[2] 黄丽晶[2] 

机构地区：[1]广州市胸科医院外科,广东广州510095 [2]广州市胸科医院呼吸病研究所,广东广州510095 [3]广州市胸科医院内科,广东广州510095

出　　处：《现代生物医学进展》2010年第10期1892-1894,共3页Progress in Modern Biomedicine

基　　金：广东省卫生厅资助科研项目(A2008554)

摘　　要：目的:比较分析空洞型肺结核组织样本与痰样本结核分枝杆菌培养的阳性率及检测的一致性,并基于rpoB基因及KatG基因突变检测在诊断耐多药结核分枝杆菌中的重要作用,探讨直接应用肺结核组织样本进行结核分枝杆菌耐药性检测的临床意义。方法:对肺结核实施肺部手术患者,在术前和术后分别进行痰及手术切除组织样本利用BacT ALERT 3D快速培养系统进行分枝杆菌培养,对培养阳性者进行菌种鉴定,对结核分枝杆菌分离株应用绝对浓度法进行异烟肼(H)、利福平(R)药物敏感性测定,比较分析二者的一致性;同时应用特异性引物对手术切除组织样本直接进行分枝杆菌rpoB、KatG基因扩增,并与其培养结果进行比较。结果:62例进入分析的病例中,术前痰培养阳性22例,阳性率35.5%(22/62),药敏试验耐药18例,耐异烟肼16例,耐利福平13例,二者均耐药12例,术中所取结核组织培养阳性34例,阳性率55%(34/62),耐药30例,耐H26例,耐R20例,二者均耐药17例,术中所取结核组织耐药基因检测与INH耐药性相关的KatG基因突变表现有22例,与RFP的耐药性相关的rpoB基因突变18例。二者同时有突变表现的16例。所有培养为结核敏感菌的组织检测中,无上述二基因突变表现。在相关的rpoB基因突变18例病例中有16例同时表现出KatG基因突(89%,16/18)。结论:厚壁空洞肺结核患者肺部手术切除组织样本病原学检测与痰样本间具有较好的一致性,且结核菌培养的阳性率明显高于普通痰培养。同时可直接应用组织样本进行结核分枝杆菌rpoB基因、KatG基因检测,可更加及时、准确地用于指导临床术后抗结核的药物治疗。Objectives：To compare the cultured positive rates of Mycobacterium tuberculosis and their detective consistency between the tissue samples of cavitary pulmonary tuberculosis and sputum samples,and to probe clinical significance of the detection of multidrug-resistant Mycobacterium tuberculosis by directly using the tissue samples of pulmonary tuberculosis based on rpoB gene and the KatG gene mutation detection in the important role of diagnosing multidrug-resistant Mycobacterium tuberculosis.Methods：Mycobacterium in the preoperative and postoperative sputum samples and the tissue samples resected by surgery from the patients with pulmonary tuberculosis were cultured by the rapid bacterial culture system of BacT ALERT 3D,cultured positive strains of which were identified in strains.The drug sensitivities of isoniazid（H） and rifampicin（R） in the isolated strains of Mycobacterium tuberculosis were respectively tested by the absolute concentration method,in which the consistency between the sputum samples and the tissue samples were compared.The gene fragments of Mycobacterium rpoB and KatG in the resected tissue samples were amplified by the specific primers,the results of which were compared with those of the culture.Results：In 62 cases,there were 22 cases of preoperative sputum culture positive,in which the positive rate was 35.5%（22/62）.The drug sensitive test showed 18 cases of drug resistance,16 only resistance to isoniazid,13 only resistance to rifampin,12 resistance to both isoniazid and rifampin.There were 34 cases of culture-positive of Mycobacterium tuberculosis in 62 cases of tuberculose focus,showing the positive rate of 55%（34/62）,30 cases of drug resistance,26 only resistance to isoniazid,20 only resistance to rifampin,17 resistance to both isoniazid and rifampin.22 cases of katG gene mutations related INH resistance,18 cases of rpoB gene mutations related RFP resistance and 16 cases of the mutation of both genes were found by the drug resistance gene testing in 62 cases of the tu

关 键 词：结核/肺 手术 RPOB基因 KATG基因 耐多药 

分 类 号：R521[医药卫生—内科学]