微孔板法在一线抗结核药物敏感性试验中的应用价值

doi:10.3969/j.issn.1000-6621.2020.04.014

摘要/Abstract

摘要：

目的分析微孔板法在一线抗结核药物药物敏感性试验(简称“药敏试验”)中的应用价值。方法收集2019年1—6月西安市胸科医院经BACTEC MGIT 960液体培养(简称MGIT 960法)培养阳性的1086份标本,选择其中经菌种鉴定为结核分枝杆菌且同时对链霉素、异烟肼、利福平、乙胺丁醇行MGIT 960 法和微孔板法药敏试验的331份菌液,分析两种试验方法的检测情况;并以MGIT 960法为标准,评估微孔板法药敏试验的检测效能,对结果不一致的菌液以熔解曲线法检测耐药基因予以核实。结果以MGIT 960法为标准,微孔板法对链霉素、异烟肼、利福平、乙胺丁醇等4种药品的敏感度、特异度、Kappa值分别为88.7%(63/71)、100.0%(260/260)、0.93,93.9%(77/82)、98.8%(246/249)、0.94,93.8%(45/48)、99.6%(282/283)、0.95,66.7%(14/21)、99.0%(307/310)、0.72。微孔板法与MGIT 960法检测4种药品耐药性结果不一致者分别为链霉素10份、异烟肼8份、利福平6份、乙胺丁醇16份。经熔解曲线耐药基因分析后,结果显示微孔板法为中敏、MGIT 960法为耐药者10份(3.0%,10/331),耐药基因检测结果均为突变,分别为链霉素2份、利福平2份、乙胺丁醇6份;而微孔板法为中敏、MGIT 960法为敏感仅乙胺丁醇1份(0.3%,1/331),耐药基因检测结果为突变;微孔板法为敏感、MGIT 960法为耐药者23份(6.9%,23/331),耐药基因检测结果为野生型10份(分别为链霉素6份、异烟肼4份)、突变型13份(分别为链霉素2份、异烟肼1份、利福平3份、乙胺丁醇7份);微孔板法为耐药、MGIT 960法为敏感者6份(1.8%,6/331),耐药基因检测结果为野生型3份(分别为异烟肼、利福平、乙胺丁醇各1份)、突变型3份(分别为异烟肼2份、乙胺丁醇1份)。结论微孔板法对一线抗结核药物的耐药性检测操作简便,与MGIT 960法一致性高,且可检测出低浓度耐药情况,可作为临床选择药物及使用剂量时的参考。

关键词: 分枝杆菌,结核, 结核,抗多种药物性, 微生物敏感性试验, 对比研究

Abstract:

Objective To explore the value of Micropore-plate method (Microdrug sensitivity, MicroDST) test in detecting sensitivity of first-line anti-tuberculosis drugs. Methods A total of 1086 specimens cultured positively by the MGIT 960 method were collected from Xi’an Chest Hospital from January to June 2019, and 331 bacterial liquids were selected which were identified as Mycobacterium tuberculosis and were tested for drug snensitivity by MicroDST and MGIT 960 method. Results of the two methods were analyzed and the detection efficiency of was evaluated with the gold standard-MGIT 960 test. Melting curve method were used to verify the resistant genotypes when test results of bacterial liquids were different. Results Based on MGIT 960 test, the sensitivity, specificity and Kappa values of MicroDST for streptomycin, isoniazid, rifampicin and ethambutol were 88.7% (63/71), 100.0% (260/260), 0.93; 93.9% (77/82), 98.8% (246/249), 0.94; 93.8% (45/48), 99.6% (282/283), 0.95; and 66.7% (14/21), 99.0% (307/310), 0.72 respectively. Among the inconsistent results of the four drug resistance tests using the MicroDST method and the MGIT 960 method, there were 10 for streptomycin, 8 for isoniazid, 6 for rifampicin, and 16 for ethambutol. In the 10 specimens (3.0%, 10/331) which were medium sensitive for MicroDST and resistant for MGIT 960 test, drug resistance gene were mutant (2 for streptomycin, 2 for rifampicin, and 6 for ethambutol). Only one specimen (0.3%, 1/331) was medium sensitive for ethambutol in MicroDST and sensitive in MGIT 960 test, the drug resistance gene was mutant. In the 23 specimens (6.9%, 23/331) which were sensitive in MicroDST and resistant in MGIT 960 test, there were 10 specimens which drug resistance gene were wild type (6 for streptomycin and 4 for isoniazid) and 13 specimens which drug resistance gene were mutant (2 for streptomycin, 1 for isoniazid, 3 for rifampicin, and 7 for ethambutol). In the 6 specimens (1.8%, 6/331) which were resistant in MicroDST and sensitive in MGIT 960 test, there were 3 specimens which drug resistance gene were wild type (1 for isoniazid, 1 for rifampicin, and 1 for ethambutol, respectively) and 3 specimen which drug resistance gene were mutant (2 for isoniazid and 1 for ethambutol). Conclusion The MicroDST method is simple and convenient for the detection of first-line anti-tuberculosis drugs. It is highly consistent with the MGIT 960 method and can detect drug resistance at low concentrations, which can provide a reference for clinical drug dosage selection.

Key words: Mycobacterium tuberculosis, Tuberculosis,multidrug-resistant, Microbial sensitivity tests, Comparative study

杨翰,杨静芬,伍浩,崔晓利,党丽云. 微孔板法在一线抗结核药物敏感性试验中的应用价值[J]. 中国防痨杂志, 2020, 42(4): 380-384. doi: 10.3969/j.issn.1000-6621.2020.04.014

YANG Han,YANG Jing-fen,WU Hao,CUI Xiao-li,DANG Li-yun. Value of MicroDST test in detecting sensitivity of first-line anti-tuberculosis drugs[J]. Chinese Journal of Antituberculosis, 2020, 42(4): 380-384. doi: 10.3969/j.issn.1000-6621.2020.04.014

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