A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis
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 imped...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-01-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.796916/full |
| _version_ | 1819283598438563840 |
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| author | Peng Liu Xinjie Wang Juan Liang Qian Dong Jinping Zhang Dongxin Liu Shuai Wang Jing Bi Wenqi Liu Zhaoqin Wang Liang Chen Lei Liu Xingxu Huang Guoliang Zhang |
| author_facet | Peng Liu Xinjie Wang Juan Liang Qian Dong Jinping Zhang Dongxin Liu Shuai Wang Jing Bi Wenqi Liu Zhaoqin Wang Liang Chen Lei Liu Xingxu Huang Guoliang Zhang |
| author_sort | Peng Liu |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-24T01:34:02Z |
| format | Article |
| id | doaj.art-7c5d5bd0f2ae41cd9b5e24124e8a46af |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-24T01:34:02Z |
| publishDate | 2022-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-7c5d5bd0f2ae41cd9b5e24124e8a46af2022-12-21T17:22:16ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-01-011210.3389/fmicb.2021.796916796916A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosisPeng Liu0Xinjie Wang1Juan Liang2Qian Dong3Jinping Zhang4Dongxin Liu5Shuai Wang6Jing Bi7Wenqi Liu8Zhaoqin Wang9Liang Chen10Lei Liu11Xingxu Huang12Guoliang Zhang13National Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaShenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, ChinaThe Biomedical Translational Research Institute, Faculty of Medical Science, Jinan University, Guangzhou, ChinaDepartment of Laboratory Medicine, Zhongshan Hospital of Sun Yat-sen University, Zhongshan, ChinaIntensive Care Unit, Liyuan Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, ChinaNational Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaNational Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaNational Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaNational Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaNational Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaGuangdong Center for Tuberculosis Control, Guangzhou, ChinaNational Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaSchool of Life Science and Technology, ShanghaiTech University, Shanghai, ChinaNational Clinical Research Center for Infectious Diseases, Guangdong Provincial Clinical Research Center for Tuberculosis, Shenzhen Third People’s Hospital, Southern University of Science and Technology, Shenzhen, ChinaDrug-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.https://www.frontiersin.org/articles/10.3389/fmicb.2021.796916/fullstreptomycindrug resistancetuberculosisCRiSPR/Casrecombinase polymerase amplification |
| spellingShingle | Peng Liu Xinjie Wang Juan Liang Qian Dong Jinping Zhang Dongxin Liu Shuai Wang Jing Bi Wenqi Liu Zhaoqin Wang Liang Chen Lei Liu Xingxu Huang Guoliang Zhang A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis Frontiers in Microbiology streptomycin drug resistance tuberculosis CRiSPR/Cas recombinase polymerase amplification |
| title | A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis |
| title_full | A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis |
| title_fullStr | A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis |
| title_full_unstemmed | A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis |
| title_short | A Recombinase Polymerase Amplification-Coupled Cas12a Mutant-Based Module for Efficient Detection of Streptomycin-Resistant Mutations in Mycobacterium tuberculosis |
| title_sort | recombinase polymerase amplification coupled cas12a mutant based module for efficient detection of streptomycin resistant mutations in mycobacterium tuberculosis |
| topic | streptomycin drug resistance tuberculosis CRiSPR/Cas recombinase polymerase amplification |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.796916/full |
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