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

文献类型： 外文期刊

作者： Liu, Peng 1 ; Wang, Xinjie 2 ; Liang, Juan 3 ; Dong, Qian 4 ; Zhang, Jinping 5 ; Liu, Dongxin 1 ; Wang, Shuai 1 ; Bi, Jing 1 ; Liu, Wenqi 1 ; Wang, Zhaoqin 1 ; Chen, Liang 6 ; Liu, Lei 1 ; Huang, Xingxu 7 ; Zhang, Guoliang 1 ;

作者机构： 1.Southern Univ Sci & Technol, Shenzhen Peoples Hosp 3, Guangdong Prov Clin Res Ctr TB, Natl Clin Res Ctr Infect Dis, Shenzhen, Peoples R China

2.Chinese Acad Agr Sci, Agr Genom Inst Shenzhen, Guangdong Lab Lingnan Modern Agr, Shenzhen Branch,Genome Anal Lab,Minist Agr & Rura, Shenzhen, Peoples R China

3.Jinan Univ, Biomed Translat Res Inst, Fac Med Sci, Guangzhou, Peoples R China

4.Sun Yat Sen Univ, Zhongshan Hosp, Dept Lab Med, Zhongshan, Peoples R China

5.Huazhong Univ Sci & Technol, Liyuan Hosp, Tongji Med Coll, Intens Care Unit, Wuhan, Peoples R China

6.Guangdong Ctr TB Control, Guangzhou, Peoples R China

7.Shanghai Tech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China

关键词： streptomycin; drug resistance; tuberculosis; CRiSPR/Cas; recombinase polymerase amplification

期刊名称：FRONTIERS IN MICROBIOLOGY （ 影响因子：6.064； 五年影响因子：6.843 ）

ISSN：

年卷期： 2022 年 12 卷

页码：

收录情况： SCI

摘要： 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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