多色巢式实时荧光定量聚合酶链式反应技术为初始方法联合线性探针及表型药敏试验的二阶段流程检测耐药结核病的临床实践

赵爱兰; 代小伟; 余琴; 张爱洁; 夏辉; 俞南

doi:10.3784/jbjc.202312070645

多色巢式实时荧光定量聚合酶链式反应技术为初始方法联合线性探针及表型药敏试验的二阶段流程检测耐药结核病的临床实践

Clinical practice of two-stage drug-resistant tuberculosis detection: multi-color nested real-time fluorescence quantitative polymerase chain reaction as initial test combined with line probe assay and phenotypic drug susceptibility test

摘要

摘要:
目的评估以多色巢式实时荧光定量聚合酶链式反应技术（GeneXpert）为初始检测方法联合线性探针及表型药敏试验的二阶段检测流程在临床使用时对利福平耐药结核病的检出效果。
方法回顾性收集并分析2019—2022年北京市朝阳区结核病门诊部登记的645例病原学阳性肺结核患者基本信息及实验室检测结果。参照《利福平耐药肺结核诊断流程》解读，将GeneXpert作为初始第一阶段利福平耐药检出方法，然后对未接受GeneXpert检测或检测结果显示结核分枝杆菌阴性或利福平耐药不明确的患者标本开展培养，并对培养阳性菌株开展线性探针耐多药检测和微孔板法进一步进行利福平耐药检测的第二阶段检测流程。对二阶段流程中检出的利福平耐药菌株进行利福平、异烟肼、左氧氟沙星、莫西沙星4种药物比例法药敏试验。分析二阶段检测流程对利福平耐药检出情况及利福平耐药患者对其他药物的耐药谱。
结果 645例病原学阳性患者的利福平耐药检测率为95.97%（619/645），利福平耐药检出比例为7.59%（47/619）。其中第一阶段GeneXpert法检出87.23%（41/47）利福平耐药患者，而第二阶段线性探针和微孔板法共检出12.77%（6/47）利福平耐药患者。 47例利福平耐药病例中37例培养阳性，经4种药物比例法药敏检测显示，7例为单耐利福平，26例为同时耐利福平和异烟肼，2例为准广泛耐药。
结论以GeneXpert为初始检测方法联合后续培养及线性探针和/或表型药敏试验的二阶段检测流程有助于检出更多利福平耐药患者。

Abstract:
Objective To evaluate the performance of the two-stage rifampin resistance tuberculosis (TB) detection with multi-color nested real-time fluorescence quantitative polymerase chain reaction (GeneXpert) as the initial test combined with line probe assay and phenotypic drug susceptibility test.
Methods Basic information and laboratory test results of 645 pathogen positive pulmonary TB patients registered in our hospital from 2019 to 2022 were collected. According to the “rifampicin-resistant pulmonary tuberculosis diagnostic algorithm”, GeneXpert was used as the initial test for rifampin resistance detection. Then, patients who did received no GeneXpert test or had negative GeneXpert results of Mycobacterium tuberculosis or unknown rifampicin resistance were provided solid culture, then line probe multidrug-resistance assay or broth microdillution test were used for the positive isolates as the second rifampin resistance detection test. Proportion drug susceptibility test to rifampin, isoniazid, levofloxacin, and moxifloxacin were conducted for the rifampicin resistant strains detected in the two-stage rifampin resistance detections. Rifampin resistance detected in the two-stage detections and overall drug resistance patterns of rifampin resistant TB cases were analyzed.
Results In 645 pathogen positive TB patients. 619 received rifampicin resistance detection (95.97%), and the rifampicin resistance rate was 7.59% (47/619). The first stage test contributed to the detection of 87.23% (41/47) of all rifampin resistant cases, while the second stage test contributed to the detection of 12.77% (6/47) of all rifampin resistant TB cases. In the 47 rifampicin resistant TB cases, 37 were cultured positive. Of the 37 cases, 7 were found to be mono rifampin resistant, 26 were resistant to both rifampicin and isoniazid, and 2 were found to be pre-extensive drug resistant indicated by 4 proportion drug susceptibility tests.
Conclusion The two-stage rifampin resistant TB detection with GeneXpert as the initial test combined with culture, line probe assay, and/or phenotypic drug sensitivity test can facilitate the diagnosis of rifampin resistant TB cases.

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