两种分子药物敏感性试验与表型药物敏感性试验检测结核分枝杆菌耐药性的效能比较

doi:10.19982/j.issn.1000-6621.20220259

摘要/Abstract

摘要：

目的:评价基因芯片法和荧光定量PCR探针熔解曲线法直接检测结核病患者临床标本中结核分枝杆菌(MTB)耐药性的价值。方法:选取解放军总医院第八医学中心结核病医学部住院结核病患者临床标本44份(例),以传统药物敏感性试验(简称“药敏试验”)绝对浓度法为对照,应用基因芯片法直接检测临床标本中MTB对利福平、异烟肼的耐药性,以及采用荧光定量PCR探针熔解曲线法直接检测临床标本中MTB对利福平、异烟肼、左氧氟沙星的耐药性,评价这两种分子药敏试验方法的诊断效能。结果:以传统药敏试验绝对浓度法检测结果为标准,应用基因芯片法检测44份(例)临床标本,MTB对利福平和异烟肼耐药性的敏感度为87.5%(14/16)和85.7%(12/14),特异度为96.4%(27/28)和96.7%(29/30),准确度为93.2%(41/44)和93.2%(41/44),Kappa值为0.851和0.758。应用荧光定量PCR探针熔解曲线法检测44份(例)临床标本,MTB对利福平、异烟肼和左氧氟沙星耐药性的敏感度为87.5%(14/16)、85.7%(12/14)和85.0%(17/20),特异度为89.3%(25/28)、83.3%(25/30)和100.0%(24/24),准确度为88.6%(39/44)、84.1%(37/44)和93.2%(41/44),Kappa值为0.840、0.653和0.861。结论:基因芯片法和荧光定量PCR探针熔解曲线法直接检测临床标本中MTB对利福平、异烟肼、左氧氟沙星的耐药性与绝对浓度法均有较高的一致性,可作为临床制定化疗方案的参考依据。

关键词: 分枝杆菌,结核, 抗药性, 分子诊断技术, 聚合酶链反应, 对比敏感度

Abstract:

Objective:To evaluate the value of the gene chip method and fluorescence quantitative PCR-probe melting curve method in the direct detection of Mycobacterium tuberculosis (MTB) drug resistance in clinical specimens of tuberculosis patients. Methods:Forty-four clinical specimens of hospitalized tuberculosis patients in the Senior Department of Tuberculosis, the 8th Medical Center of Chinese PLA General Hospital were selected. Using the absolute concentration method of the traditional drug susceptibility test as the control, the drug resistance of MTB to rifampicin and isoniazid in clinical samples was directly detected by the gene chip method, and the drug resistance of MTB to rifampicin, isoniazid, and levofloxacin in the clinical specimens was directly detected by fluorescence quantitative PCR-probe melting curve method, to evaluate the sensitivity, specificity, and consistency of these two molecular drug susceptibility test methods. Results:The traditional drug susceptibility test was used as the standard, the sensitivity, specificity, consistency, and Kappa value of MTB to rifampicin resistance in 44 clinical samples directly detected by the gene chip method were 87.5% (14/16), 96.4% (27/28), 93.2% (41/44) and 0.851, respectively; for the isoniazid resistance were 85.7% (12/14), 96.7% (29/30), 93.2% (41/44) and 0.758, respectively. The sensitivity, specificity, consistency, and Kappa value of MTB to rifampin resistance in 44 clinical samples directly detected by the probe melting curve method were 87.5% (14/16), 89.3% (25/28), 88.6% (39/44), and 0.840, respectively; for isoniazid resistance were 85.7% (12/14), 83.3% (25/30), 84.1% (37/44), and 0.653, respectively; for the levofloxacin resistance were 85.0% (17/20), 100.0% (24/24), 93.2% (41/44), and 0.861, respectively. Conclusion:The gene chip method and probe melthing curve method have a high consistency with the absolute concentration method in detecting the drug resistant of MTB to rifampicin, isoniazid (and levofloxain) in clinical specimens,which can be used as a reference for clinical chemotherapy.

Key words: Mycobacterium tuberculosis, Drug resistance, Molecular diagnostic techniques, Polymerase chain reaction, Contrast sensitivity

中图分类号:

R52
U472.9

孙雯娜, 张俊仙, 张秀爽, 王晓朦, 林文, 梁艳, 王杰, 吴雪琼. 两种分子药物敏感性试验与表型药物敏感性试验检测结核分枝杆菌耐药性的效能比较[J]. 中国防痨杂志, 2022, 44(10): 1016-1021. doi: 10.19982/j.issn.1000-6621.20220259

Sun Wenna, Zhang Junxian, Zhang Xiushuang, Wang Xiaomeng, Lin Wen, Liang Yan, Wang Jie, Wu Xueqiong. Efficacy comparison of two molecular drug sensitivity methods and phenotypic drug sensitivity testindetecting drug resistance of Mycobacterium tuberculosis[J]. Chinese Journal of Antituberculosis, 2022, 44(10): 1016-1021. doi: 10.19982/j.issn.1000-6621.20220259

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传统药敏试验绝对浓度法	芯片法(例)		合计 (例)	敏感度 (%, 95%CI)	特异度 (%,95%CI)	准确度 (%,95%CI)	Kappa值 (95%CI)
传统药敏试验绝对浓度法	耐药	敏感	合计 (例)	敏感度 (%, 95%CI)	特异度 (%,95%CI)	准确度 (%,95%CI)	Kappa值 (95%CI)
利福平				87.5(60.4~97.8)	96.4(79.8~99.8)	93.2(81.3~98.6)	0.851(0.688~1.000)
耐药	14	2	16
敏感	1	27	28
合计	15	29	44
异烟肼				85.7(56.2~97.5)	96.7(81.0~99.8)	93.2(81.3~98.6)	0.758(0.588~0.958)
耐药	12	2	14
敏感	1	29	30
合计	13	31	44

传统药敏试验绝对浓度法	熔解曲线法
传统药敏试验绝对浓度法	耐药	敏感
高耐(17)	gyrA 88~94(16)	1
低耐(3)	gyrA 88~94(1)	2
敏感(24)	0	24
合计	17	27