Scholarlink

Background <p>Rapidly detecting drug-resistant <i>Mycobacterium tuberculosis</i> (MTB) is essential for effective tuberculosis (TB) treatment. We have developed a one-tube nested fluorescence melting curve assay (OTN-FMCA) that allows rapid and simultaneous detection of resistance to rifampicin, isoniazid, fluoroquinolones, and bedaquiline.</p> Results <p>The OTN-FMCA uses nested PCR and locked nucleic acid (LNA)-modified probes to identify 44 mutations in six key genes (<i>rpoB</i>,<i> katG</i>,<i> inhA</i>,<i> gyrA</i>,<i> gyrB</i>, and <i>atpE</i>), achieving a detection limit of 5 copies per microliter and identifying up to 10% heterogeneous resistance. When tested on 198 culture-positive isolates, OTN-FMCA demonstrated high concordance with microarray susceptibility testing, with 100% sensitivity and 96.89% specificity. Further validation with 518 sputum samples confirmed its ability to classify patients into different drug resistance categories, including DS-TB, RR-TB, MDR-TB, pre-XDR-TB, and XDR-TB. OTN-FMCA offers a faster, more affordable, and more accessible alternative to current comprehensive drug resistance profiling methods, especially in resource-limited settings.</p> Conclusions <p>OTN-FMCA is a promising tool for the rapid and accurate detection of drug resistance in MTB, which could improve the management of drug-resistant TB.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

A novel one-tube nested fluorescence melting curve analysis for rapid detection of drug-resistant Mycobacterium tuberculosis

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年12月20日

查看全文
  • Yingchao Chang,
  • Gaowen Liu,
  • Xiaofei Li,
  • Mi Zhang,
  • Tingting Diao,
  • Cuixian Yang,
  • Xinlin Wu,
  • Yungui Tu,
  • Hongji Zhang,
  • Chao Zhou,
  • Pei Lei,
  • Li Liu,
  • Xueshan Xia,
  • Yue Feng

摘要

Background

Rapidly detecting drug-resistant Mycobacterium tuberculosis (MTB) is essential for effective tuberculosis (TB) treatment. We have developed a one-tube nested fluorescence melting curve assay (OTN-FMCA) that allows rapid and simultaneous detection of resistance to rifampicin, isoniazid, fluoroquinolones, and bedaquiline.

Results

The OTN-FMCA uses nested PCR and locked nucleic acid (LNA)-modified probes to identify 44 mutations in six key genes (rpoB, katG, inhA, gyrA, gyrB, and atpE), achieving a detection limit of 5 copies per microliter and identifying up to 10% heterogeneous resistance. When tested on 198 culture-positive isolates, OTN-FMCA demonstrated high concordance with microarray susceptibility testing, with 100% sensitivity and 96.89% specificity. Further validation with 518 sputum samples confirmed its ability to classify patients into different drug resistance categories, including DS-TB, RR-TB, MDR-TB, pre-XDR-TB, and XDR-TB. OTN-FMCA offers a faster, more affordable, and more accessible alternative to current comprehensive drug resistance profiling methods, especially in resource-limited settings.

Conclusions

OTN-FMCA is a promising tool for the rapid and accurate detection of drug resistance in MTB, which could improve the management of drug-resistant TB.