| 林翀 何海武 何晶 周云 林明冠 钟业腾 陈少文 廉芳 苏应仙.高分辨率熔解曲线分析技术检测结核分枝杆菌的耐药性[J].现代生物医学进展英文版,2016,16(17):3256-3260. |
| 高分辨率熔解曲线分析技术检测结核分枝杆菌的耐药性 |
| Detection of Multidrug-resistant Mycobacterium Tuberculosis by HighResolution Melting Analysis of Multidrug-resistant by HighResolution Melting Analysis |
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| DOI： |
| 中文关键词: 高分辨率熔解曲线 结核 耐药 特异性 灵敏度 |
| 英文关键词: High resolution melting Tuberculosis Drug-resistance Specificity Sensitivity |
| 基金项目:海南省卫生厅医学科研立项课项（琼卫科教〔2014〕51 号）
琼卫2013 资助-058 号 |
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| 中文摘要: |
| 目的：探讨高分辨率熔解曲线分析(High resolution melting, HRM)技术检测结核分枝杆菌耐药突变位点的可行性。方法：对
218 株结核分枝杆菌进行利福平(RFP)和异烟肼(INH)的药物敏感性测定，并进行耐药基因位点的PCR 扩增和测序，同时采用
HRM方法检测RFP 和INH耐药基因位点情况，分析HRM的敏感性和特异性。结果：218株结核分枝杆菌药敏试验结果显示，有
106株(48.6%)对RFP 耐药，100 株(45.9%)对INH 耐药，81 株(37.4%)对RFP和INH 均耐药。测序发现，101 株(46.3%)存在RFP耐
药基因的突变，107 株(49.1%)存在INH耐药基因的突变。HRM 检测结果显示，100 株(45.9%)存在RFP 耐药基因的突变，103 株
(47.2%)存在INH耐药基因的突变。分别以药敏试验和测序结果为标准，HRM检测RFP耐药的敏感性为94.3%(100/106)和99.0%
(100/101)；特异性为97.3%(109/112)和100%(117/117)；INH 耐药的敏感性为97.0%(97/100)和98.1%(103/105)；特异性为97.3%
(109/112)和100%(113/113)。结论：HRM快速检测结核分枝杆菌耐药具有较高的特异性和灵敏度，能够满足临床需求。 |
| 英文摘要: |
| Objective:To explore the feasibility in the mutation sites of Mycobacterimun tberculosis resistance by the method of
high resolution melting (HRM).Methods:218 strains of Mycobacterium tuberculosis were collected to complete the drug sensibilities test
of rifampin (RFP) and isoniazid (INH). The drug resistant sites were amplified by PCR and sequenced. HRM method was performed to
detect the mutation sites of RFP and INH which leads to mycobacterium tuberculosis resistance, and to analyze the sensitivity and specificity
of HRM method.Results:There were 106 (48.6%) RFP resistant strains, 100 (45.9%) INH resistant strains, 81 (37.4%) both RFP
and INH resistant strains in drug sensibilities of 218 strains mycobacterium tuberculosis. We found 101 (46.3%) and 100 (45.9%) strains
of RFP resistant gene mutation, 107 (49.1%) and 103 (47.2%) strains of INH resistant gene mutation by sequencing and HRMmethod respectively.
Compared against the results of drug sensitive test and sequencing method, for detecting RFP resistance, the sensitivity of
HRMwere 94.3% (100/106) and 99.0% (100/101), and specificity were 97.3% (109/112) and 100% (117/117). For detecting INH resistance,
the sensitivity of HRMwere 97.0% (97/100) and 98.1% (103/105), and specificity were 97.3% (109/112) and 100%(113/113).Conclusion:The detection of resistance Mycobacterium tuberculosis by HRMmethod has a higher sensitivity and specificity to meet the
clinical application. |
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