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荧光PCR熔解曲线法检测耐多药肺结核患者对左氧氟沙星和莫西沙星耐药性的效能研究
引用本文:金龙,田琦,张宝庆,邢海冬,高明霞,张秀英,王利华,张晓磊. 荧光PCR熔解曲线法检测耐多药肺结核患者对左氧氟沙星和莫西沙星耐药性的效能研究[J]. 中国防痨杂志, 2022, 44(2): 159-163. DOI: 10.19982/j.issn.1000-6621.20210488
作者姓名:金龙  田琦  张宝庆  邢海冬  高明霞  张秀英  王利华  张晓磊
作者单位:1.黑龙江省传染病防治院结核内科,哈尔滨 150500;2.黑龙江省传染病防治院结核病实验室,哈尔滨 150500
基金项目:黑龙江省卫生计生委科研课题(2018-586)。
摘    要:目的: 探究荧光PCR熔解曲线法快速检测耐多药肺结核患者对左氧氟沙星(Lfx)和莫西沙星(Mfx)耐药性的临床价值。 方法: 以2018年11月至2019年12月在黑龙江省传染病防治院采用BACTEC MGIT 960全自动分枝杆菌培养及比例法药物敏感性试验(简称“药敏试验”)确诊为耐多药肺结核的195例患者为研究对象,同时经荧光PCR熔解曲线法进行筛查,通过与药敏试验结果进行对比,分析荧光PCR熔解曲线法检测结核分枝杆菌对Lfx和Mfx耐药性的准确性。结果: 195株耐多药结核分枝杆菌经比例法药敏试验检测,对Lfx和Mfx耐药率分别为46.15%(90/195)和48.72%(95/195);以比例法药敏试验结果为标准,荧光PCR熔解曲线法检测结核分枝杆菌对Lfx和Mfx耐药性的敏感度分别为95.56%(86/90)和95.79%(91/95),特异度分别为73.33%(77/105)和77.00%(77/100),一致率分别为83.59%(163/195)和86.15%(168/195),Kappa值分别为0.83和0.85。 结论: 荧光PCR熔解曲线法检测耐多药肺结核患者对Lfx和Mfx耐药的结果与比例法药敏试验结果高度一致,适用于筛查耐多药肺结核患者对Lfx和Mfx的耐药性。

关 键 词:结核    结核  抗多种药物性  喹诺酮类  分子诊断技术  诊断敏感试验  
收稿时间:2021-08-13

Research of effectiveness of fluorescence PCR melting curve assay in detecting drug resistance of levofloxacin and moxifloxacin in patients with multidrug-resistant pulmonary tuberculosis
JIN Long,TIAN Qi,ZHANG Bao-qing,XING Hai-dong,GAO Ming-xia,ZHANG Xiu-ying,WANG Li-hua,ZHANG Xiao-lei. Research of effectiveness of fluorescence PCR melting curve assay in detecting drug resistance of levofloxacin and moxifloxacin in patients with multidrug-resistant pulmonary tuberculosis[J]. The Journal of The Chinese Antituberculosis Association, 2022, 44(2): 159-163. DOI: 10.19982/j.issn.1000-6621.20210488
Authors:JIN Long  TIAN Qi  ZHANG Bao-qing  XING Hai-dong  GAO Ming-xia  ZHANG Xiu-ying  WANG Li-hua  ZHANG Xiao-lei
Affiliation:1.Department of Tuberculosis, Infectious Disease Hospital of Heilongjiang Province, Harbin 150500, China;2.Tuberculosis Laboratory, Infectious Disease Hospital of Heilongjiang Province, Harbin 150500, China
Abstract:Objective: To explore the clinical value of fluorescence PCR melting curve assay for rapid detection of levofloxacin (Lfx) and moxifloxacin (Mfx) resistance of multidrug-resistant pulmonary tuberculosis (MDR-TB) patients. Methods: From November 2018 to December 2019, 195 MDR-PTB patients from Infectious Disease Hospital of Heilongjiang Province were selected as subjects. All the patients were screened by fluorescence PCR melting curve assay and confirmed by the automatic BACTEC MGIT 960 Mycobacteria Culture System and proportion method of drug sensitivity test (DST). The accuracy of fluorescence PCR melting curve assay for detecting drug resistance to Lfx and Mfx of Mycobacterium tuberculosis (MTB) was analyzed. Results: Using the proportion method, among the 195 MDR-MTB strains, the drug resistance rates of Lfx and Mfx were 46.15% (90/195) and 48.72% (95/195), respectively. Using the above results as standard, the sensitivity, specificity, consistency and Kappa value of Lfx resistance of MTB were 95.56% (86/90), 73.33% (77/105), 83.59% (163/195) and 0.83; and the sensitivity, specificity, consistency and Kappa value of Mfx resistance of MTB were 95.79% (91/95), 77.00% (77/100), 86.15% (168/195) and 0.85 by fluorescence PCR melting curve analysis. Conclusion: The results of fluorescence PCR melting curve assay in detecting the resistance of MDR-TB to Lfx and Mfx were highly consistent with the results of proportion method of DST. Fluorescence PCR melting curve analysis assay is suitable for screening the drug resistance of MDR-TB to Lfx and Mfx.
Keywords:Tuberculosis,pulmonary  Tuberculosis,multidrug-resistant  Quinolones  Molecular diagnostic techniques  Diagnostic self evaluation
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