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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。
OBJECTIVE: To compare and analyze the positive rate and the consistency of detection of Mycobacterium tuberculosis in sputum of tuberculosis and tuberculosis, and to study the important role of rpoB gene and KatG gene mutation in diagnosis of multidrug-resistant Mycobacterium tuberculosis Clinical Significance of Direct Application of Pulmonary Tuberculosis Tissues to Mycobacterium Tuberculosis Drug Resistance Test. Methods: Patients with pulmonary tuberculosis were treated with sputum and surgical resection respectively before and after operation. BacT ALERT 3D rapid culture system was used to culture mycobacteria. The positive cultures were identified, Bacillus isolate absolute concentration method for determination of isoniazid (H), rifampin (R) drug sensitivity, comparative analysis of the consistency between the two; at the same time using specific primers for surgical removal of tissue samples direct mycobacterial rpoB , KatG gene amplification, and its culture results were compared. Results: Of the 62 cases who were included in the analysis, 22 were positive sputum cultures (35.5%, 22/62), 18 were resistant to antimicrobial susceptibility test, 16 were resistant to isoniazid, 13 were resistant to rifampicin, Both were drug-resistant in 12 cases, during which tuberculosis tissue culture positive 34 cases, the positive rate was 55% (34/62), resistance in 30 cases, resistant to H26 cases, resistant to R20 cases, both 17 cases of drug resistance, There were 22 cases of KatG gene mutation associated with INH resistance and 18 cases of rpoB gene mutation associated with drug resistance of RFP. Both of them showed mutation in 16 cases at the same time. Tissue testing of all those cultivated as sensitive tuberculosis did not show the above two mutations. Of the 18 cases of related rpoB mutations, 16 also showed KatG gene mutations (89%, 16/18). Conclusion: There is a good agreement between etiological detection and sputum samples in patients with thick-walled hollow pulmonary tuberculosis. The positive rate of TB culture was significantly higher than that of common sputum culture. At the same time, the samples of Mycobacterium tuberculosis rpoB gene and KatG gene can be directly applied to the tissue samples, which can be more timely and accurately used to guide the clinical postoperative anti-TB drug treatment.