4.6 Article

A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis

期刊

FRONTIERS IN MICROBIOLOGY
卷 12, 期 -, 页码 -

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FRONTIERS MEDIA SA
DOI: 10.3389/fmicb.2021.796916

关键词

streptomycin; drug resistance; tuberculosis; CRiSPR/Cas; recombinase polymerase amplification

资金

  1. National Key Research and Development Plan [2020YFA0907201, 2019YFC0840602, 2021YFA1300902]
  2. National Natural Science Foundation of China [81873958, 82170009]
  3. Guangdong Scientific and Technological Foundation [2019B1515120041, 2020B1111170014]
  4. Shenzhen Scientific and Technological Project [KCXFZ202002011007083]
  5. Sanming Project of Medicine in Shenzhen [SZSM201911009]

向作者/读者索取更多资源

This study developed a Cas12a RR detection system that can rapidly, simply, sensitively, and specifically detect STR-resistant TB. The system can detect target substances as low as 0.1% and the entire detection process can be completed within 60 minutes. Its sensitivity and specificity for detecting clinical STR-resistant Mycobacterium tuberculosis were both 100%.
Drug-resistant tuberculosis (TB) is a serious public health problem and threat to global TB prevention and control. Streptomycin (STR) is the earliest and classical anti-TB drug, and it is the earliest drug that generated resistance to anti-TB treatment, which limits its use in treating TB and impedes TB control efforts. The rapid, economical, and highly sensitive detection of STR-resistant TB may help reduce disease transmission and morbimortality. CRISPR/CRISPR-associated protein (Cas) is a new-generation pathogen detection method that can detect single-nucleotide polymorphisms with high sensitivity and good specificity. In this study, a Cas12a RR detection system that can recognize more non-traditional protospacer-adjacent motif-targeting sequences was developed based on Cas12a combined with recombinase polymerase amplification technology. This system detects 0.1% of the target substance, and the entire detection process can be completed within 60 min. Its sensitivity and specificity for detecting clinical STR-resistant Mycobacterium tuberculosis were both 100%. Overall, the Cas12 RR detection system provides a novel alternative for the rapid, simple, sensitive, and specific detection of STR-resistant TB, which may contribute to the prompt treatment and prevention of disease transmission in STR-resistant TB.

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