The potential of genetic testing for rapid and accurate diagnosis of drug-resistant Mycobacterium tuberculosis strains is vital for efficient treatment and reduction in dissemination. MTBDR plus assays rapidly detect mutations related to drug resistance and wild type sequences allied with susceptibility. Although these methods are promising, the examination of molecular level performance is essential for improved assay result interpretation and continued diagnostic development. Therefore this study aimed to determine novel mutations that were inhibiting wild type probe hybridization in the Line probe assay by DNA sequencing. Using data collected from Line Probe assay (GenoType MTBDRplus assay) the contribution of absent wild type probe hybridization to the detection of rifampicin resistance was assessed via comparison to a reference standard method i.e. DNA sequencing. Sequence analysis of the rpoB gene of 47 MTB resistant strains from clinical specimens showed that 37 had a single mutation, 9 had double mutations and one had triple mutations in the ropB gene. The absence of wild type probe hybridization without mutation probe hybridization was mainly the result of the failure of mutation probe hybridization and the result of the novel or rare mutations. Additional probes are necessary to be included in the Line probe assay to improve the detection of rifampicin-resistant Mycobacterium tuberculosis strains.

中文翻译：

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通过直接来自临床标本的 DNA 测序，在 MTBDR plus 测定中检测抑制利福平耐药结核分枝杆菌菌株的 rpoB 基因突变，抑制野生型探针杂交。

基因检测在快速准确诊断耐药结核分枝杆菌菌株方面的潜力对于有效治疗和减少传播至关重要。MTBDR plus 检测可快速检测与耐药性和与易感性相关的野生型序列相关的突变。尽管这些方法很有前景，但分子水平性能的检查对于改进分析结果解释和持续的诊断开发至关重要。因此，本研究旨在通过 DNA 测序确定在 Line 探针测定中抑制野生型探针杂交的新突变。使用从 Line Probe 测定（GenoType MTBDRplus 测定）收集的数据，通过与参考标准方法（即 DNA 测序）的比较来评估缺少野生型探针杂交对检测利福平抗性的贡献。对临床标本中47株MTB耐药菌株的rpoB基因序列分析显示，ropB基因37株为单突变，9株为双突变，1株为三突变。没有突变探针杂交的野生型探针杂交的缺失主要是突变探针杂交失败的结果和新的或稀有突变的结果。需要在 Line 探针检测中加入额外的探针，以改进对利福平耐药的结核分枝杆菌菌株的检测。对临床标本中47株MTB耐药菌株的rpoB基因序列分析显示，ropB基因37株为单突变，9株为双突变，1株为三突变。没有突变探针杂交的野生型探针杂交的缺失主要是突变探针杂交失败的结果和新的或稀有突变的结果。需要在 Line 探针检测中加入额外的探针，以改进对利福平耐药的结核分枝杆菌菌株的检测。对临床标本中47株MTB耐药菌株的rpoB基因序列分析显示，ropB基因37株为单突变，9株为双突变，1株为三突变。没有突变探针杂交的野生型探针杂交的缺失主要是突变探针杂交失败的结果和新的或稀有突变的结果。需要在 Line 探针检测中加入额外的探针，以改进对利福平耐药的结核分枝杆菌菌株的检测。没有突变探针杂交的野生型探针杂交的缺失主要是突变探针杂交失败的结果和新的或稀有突变的结果。需要在 Line 探针检测中加入额外的探针，以改进对利福平耐药的结核分枝杆菌菌株的检测。没有突变探针杂交的野生型探针杂交的缺失主要是突变探针杂交失败的结果和新的或稀有突变的结果。需要在 Line 探针检测中加入额外的探针，以改进对利福平耐药的结核分枝杆菌菌株的检测。