分子生物学方法在结核分枝杆菌及其耐药性检测中的价值

陈妍汶; 杨勇; 陈俊莉; 罗军; 庄利东

doi:10.13898/j.cnki.issn.1000-2200.2020.03.028

分子生物学方法在结核分枝杆菌及其耐药性检测中的价值

Value of molecular biology method in detection of Mycobacterium tuberculosis and its drug resistance

摘要

摘要:
目的探讨分子生物学方法在结核分枝杆菌（Mycobacterium tuberculosis，MTB）及其耐药性检测中的价值。
方法以MTB标准菌株H37Rν作为对照，以单耐药及耐多药的MTB行梯度浓度稀释后作为检测样本，分析Xpert MTB/RIF法、基因芯片法、改良罗氏培养法、比例法检测MTB的准确性、最低检出限及检测周期等。
结果改良罗氏培养法和Xpert MTB/RIF法检测MTB下限均为3×10² cfu/mL，基因芯片法检出下限为3×10⁴ cfu/mL；Xpert MTB/RIF法在3×10² cfu/mL时检测利福平耐药性存在假阳性现象，无法检测异烟肼的耐药性；Xpert MTB/RIF法检测周期≤2 h，而基因芯片法≤8 h，比例法和改良罗氏培养法检测周期在5~8周。
结论在MTB及其耐药性检测中分子生物学方法的检测周期优势明显，在最低检出限和准确性方面各有优势，因此，临床诊断中综合考虑病人的经济水平和病情危重程度，选择合适的检测方法。

Abstract:
ObjectiveTo explore the value of molecular biology method in the detection of Mycobacterium tuberculosis (MTB) and its drug resistance.
MethodsThe accuracy, minimum detection limit and detection period of Xpert MTB/RIF assay, gene chip assay, modified Roche culture assay and proportion assay in detecting MTB were analyzed using standard MTB strain H37Rr as control, and single-drug resistant and multi-drug resistant MTB diluted gradiently as the test sample.
ResultsThe minimum detection limit of modified Roche culture assay and Xpert MTB/RIF assay was 3×10² cfu/mL, which was 3×10⁴ cfu/mL for gene chip assay.Xpert MTB/RIF assay resulted in false-positive rifampicin resistance when the concentration was 3×10² cfu/mL, and could not detect isoniazid resistance.The detection period of Xpert MTB/RIF assay was ≤ 2 h, which was ≤ 8 h for gene chip assay, and five to eight weeks for modified Roche culture assay and proportion assay.
ConclusionsIn the detection of MTB and its drug resistance, molecular biology methods have obvious advantages in detection period, and each method has its own advantages in the minimum detection limit and accuracy.The economic level and critical degree of patients should be considered in the clinical diagnosis, so to choose appropriate detecting method.

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