线性探针技术诊断HIV/结核分枝杆菌感染者结核菌耐药性效能评价
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摘要
目的评价线性探针技术(line-probe assay, LipA)检测HIV/结核分枝杆菌(Mycobacterium tuberculosis, MTB)感染者耐药结核菌的效能,并探讨耐药结核菌基因突变特征。方法采用LipA和药物敏感(药敏)试验绝对浓度法检测HIV/MTB感染者涂阳痰标本和临床分离株中MTB对利福平(rifampicin, RFP)及异烟肼(isoniazid, INH)的耐药性,以药敏试验结果为标准评价LipA的检测效能,统计耐药MTB基因突变型。结果 LipA对涂阳痰标本和临床分离株中MTB耐RFP的检测结果与药敏试验结果完全一致,对耐INH的检测结果与药敏试验结果比较,kappa=0.969(P <0.05),耐RFP株及耐INH株的主要基因突变位点分别是rpoB S531L(80.00%)和katG 315(82.76%)。结论 LipA检测HIV/MTB感染者MTB耐RFP和INH的诊断与药敏试验结果一致,主要基因突变位点与结核病总体人群相似,适用于HIV/MTB感染者耐多药结核病的快速筛查。
Objective To evaluate the performance of line probe assay(LipA) in the diagnosis of drug-resistant Mycobacterium tuberculosis(MTB) and analyze the mutation characters of MTB in HIV/MTB co-infection patients. Methods Lip A and drug sensitivity test(DST) performed by absolute concentration method were used to detect the rifampicin(RFP) and isoniazid(INH) tolerant mutants in smear-positive sputum samples and clinical MTB isolates. The performance of LipA in the detection of drug-resistant MTB was evaluated based on the DST results, and the gene characters of MTB mutants were analyzed. Results The detection results between LipA and DST were completely identical for RFP-resistant MTB in smear-positive sputum samples and clinical MTB isolates, and showed a kappa value of 0.969(P <0.05) for INH-resistant mutants. rpo B S531L(80.00%) and kat G 315(82.76%) were the main mutation sites in RFP-and INH-resistant strains. Conclusions The performance of LipA is highly consistent with the DST in detecting RFP-and INH-resistant MTB for the HIV/MTB co-infection. The main mutation sites from the HIV/MTB co-infection are similar to those from normal tuberculosis cases. LipA is a reliable tool to detect multi-drug resistant tuberculosis for HIV/MTB co-infection rapidly.
Objective To evaluate the performance of line probe assay(LipA) in the diagnosis of drug-resistant Mycobacterium tuberculosis(MTB) and analyze the mutation characters of MTB in HIV/MTB co-infection patients. Methods Lip A and drug sensitivity test(DST) performed by absolute concentration method were used to detect the rifampicin(RFP) and isoniazid(INH) tolerant mutants in smear-positive sputum samples and clinical MTB isolates. The performance of LipA in the detection of drug-resistant MTB was evaluated based on the DST results, and the gene characters of MTB mutants were analyzed. Results The detection results between LipA and DST were completely identical for RFP-resistant MTB in smear-positive sputum samples and clinical MTB isolates, and showed a kappa value of 0.969(P <0.05) for INH-resistant mutants. rpo B S531L(80.00%) and kat G 315(82.76%) were the main mutation sites in RFP-and INH-resistant strains. Conclusions The performance of LipA is highly consistent with the DST in detecting RFP-and INH-resistant MTB for the HIV/MTB co-infection. The main mutation sites from the HIV/MTB co-infection are similar to those from normal tuberculosis cases. LipA is a reliable tool to detect multi-drug resistant tuberculosis for HIV/MTB co-infection rapidly.
引文
[1]World Health Organization. Tuberculosis fact sheet[EB/OL].[2018-03-10]. http://www. who. int/mediacentre/factsheets/fs104/en/.
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[8]汪明,袁乐永,刘乐,等. MTBDRplus方法快速检测结核分枝杆菌耐药性的研究[J].国际呼吸杂志,2015,35(21):1601-1605.
[9]Cavusoglu C, Turhan A, Akinci P, et al. Evaluation of the genotype MTBDR assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis isolates[J]. Clin Microbiol, 2006, 44(7):2338-2342.
[10]马利,茹浩浩,陈连,等.线性探针法检测云南地区结核分枝杆菌耐药的应用评价[J].临床检验志,2015,33(8):573-576.
[11]李辉,马晓光,石洁,等.线性探针法MTBDRplus用于涂阳肺结核的耐多药检测研究[J].中国卫生检验杂志,2013,23(2):380-384.
[12]张海晴,黄海滨,刘成永,等.徐州地区结核分枝杆菌基因分型及其与利福平耐药基因突变的关系[J].广东医学,2017,38(2):223-224,228.
[13]徐凯进,嵇仲,康胡海,等.浙江省12个区(县)分枝杆菌菌种鉴定及耐药相关基因特征分析[J].中国防痨杂志,2017,39(2):154-157.
[14]高旭,李静,柳清云,等.异质性耐药对结核分枝杆菌表型和基因型耐药检测结果的影响[J].中华结核与呼吸杂志,2014,34(4):260-265.
[15]MartinA,HerranzM,RuizSerranoMJ, etal.Theclonal composition of Mycobacterium tuberculosis in clinical specimens could be modified by culture[J].Tuberculosis(Edinb), 2010, 90(3):201-207.
[2]GuptaRK,LucasSB,FieldingKL, etal.Prevalenceof tuberculosis in post-mortem studies of HIV-infected adults andchildreninresource-limitedsettings:asystematic review and metaanalysis[J]. AIDS, 2015, 29(15):1987-2002.
[3]伍秋云,卢祥婵,黄爱春,等. 126例HIV/结核分枝杆菌合并感染临床观察[J].传染病信息,2015,28(2):108-111.
[4]World Health Organization. Implementing tuberculosis diagnostics.Policy framework[M]. Geneva:World Health Organization, 2015.
[5]中华医学会感染病学分会艾滋病学组.艾滋病诊疗指南(第三版)[J].中华传染病杂志,2015,33(10):577-593.
[6]中华医学会结核病学分会.肺结核诊断和治疗指南[J].中华结核和呼吸杂志,2001,24(2):70-74.
[7]赵雁林.结核病实验室标准化操作与网络建设[M].北京:人民卫生出版社,2013:69-265.
[8]汪明,袁乐永,刘乐,等. MTBDRplus方法快速检测结核分枝杆菌耐药性的研究[J].国际呼吸杂志,2015,35(21):1601-1605.
[9]Cavusoglu C, Turhan A, Akinci P, et al. Evaluation of the genotype MTBDR assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis isolates[J]. Clin Microbiol, 2006, 44(7):2338-2342.
[10]马利,茹浩浩,陈连,等.线性探针法检测云南地区结核分枝杆菌耐药的应用评价[J].临床检验志,2015,33(8):573-576.
[11]李辉,马晓光,石洁,等.线性探针法MTBDRplus用于涂阳肺结核的耐多药检测研究[J].中国卫生检验杂志,2013,23(2):380-384.
[12]张海晴,黄海滨,刘成永,等.徐州地区结核分枝杆菌基因分型及其与利福平耐药基因突变的关系[J].广东医学,2017,38(2):223-224,228.
[13]徐凯进,嵇仲,康胡海,等.浙江省12个区(县)分枝杆菌菌种鉴定及耐药相关基因特征分析[J].中国防痨杂志,2017,39(2):154-157.
[14]高旭,李静,柳清云,等.异质性耐药对结核分枝杆菌表型和基因型耐药检测结果的影响[J].中华结核与呼吸杂志,2014,34(4):260-265.
[15]MartinA,HerranzM,RuizSerranoMJ, etal.Theclonal composition of Mycobacterium tuberculosis in clinical specimens could be modified by culture[J].Tuberculosis(Edinb), 2010, 90(3):201-207.