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

Liu, Peng1; Wang, Xinjie2; Liang, Juan3; Dong, Qian4; Zhang, Jinping5; Liu, Dongxin1; Wang, Shuai1; Bi, Jing1; Liu, Wenqi1; Wang, Zhaoqin1; Chen, Liang6; Liu, Lei1; Huang, Xingxu7; Zhang, Guoliang1

2022-01-06

10.3389/fmicb.2021.796916

FRONTIERS IN MICROBIOLOGY   影响因子和分区

1664-302X

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.

streptomycin drug resistance tuberculosis CRiSPR/Cas recombinase polymerase amplification

SCI

英语

第一 ; 通讯

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

Microbiology

Microbiology

WOS:000747185500001

FRONTIERS MEDIA SA

Web of Science

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

Liu, Peng,Wang, Xinjie,Liang, Juan,et al. A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis[J]. FRONTIERS IN MICROBIOLOGY,2022,12.

Liu, Peng.,Wang, Xinjie.,Liang, Juan.,Dong, Qian.,Zhang, Jinping.,...&Zhang, Guoliang.(2022).A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis.FRONTIERS IN MICROBIOLOGY,12.

Liu, Peng,et al."A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis".FRONTIERS IN MICROBIOLOGY 12(2022).