Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplification
ObjectiveTo establish a rapid， sensitive and specific detection method for 4 diarrheal bacteria （Salmonella， Shigella， Vibrio cholera and Escherichia coli O157：H7） by the Clustered regularly interspaced short palindromic repeats associated protein 13a （CRISPR-Cas13a） combined with recombinant enzyme...
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| Format: | Article |
| Language: | zho |
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The Editorial Office of Chinese Journal of Food Hygiene
2023-03-01
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| Series: | Zhongguo shipin weisheng zazhi |
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| Online Access: | http://www.zgspws.com/zgspwszz/article/abstract/202303010 |
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| author | AN Bailin SU Xuan GUO Yue WANG Xiangxi GE Yiyue ZHU Fengcai CUI Lunbiao |
| author_facet | AN Bailin SU Xuan GUO Yue WANG Xiangxi GE Yiyue ZHU Fengcai CUI Lunbiao |
| author_sort | AN Bailin |
| collection | DOAJ |
| description | ObjectiveTo establish a rapid， sensitive and specific detection method for 4 diarrheal bacteria （Salmonella， Shigella， Vibrio cholera and Escherichia coli O157：H7） by the Clustered regularly interspaced short palindromic repeats associated protein 13a （CRISPR-Cas13a） combined with recombinant enzyme-mediated isothermal amplification （RAA）， called RAA-Cas13a.MethodsIn this study， the specific primer for RAA and CRISPR RNA （crRNA） of 4 different diarrheal bacteria were designed. The sample nucleic acids were amplified by RAA， and the amplification products were then detected with CRISPR-Cas13a. Compared with real-time quantitative polymerase chain reaction（RT-qPCR）， the sensitivity and specificity of the RAA-Cas13a method were evaluated.ResultsThe established RAA-Cas13a detection method for Shigella， Vibrio cholera and Escherichia coli O157：H7 had the detection limit of 10 copies/μL， the detection limit for Salmonella was 1 copy/μL， and each bacteria did not have cross-reaction with the other ten bacteria. Meantime， the detection of the RT-qPCR and RAA-Cas13a were highly consistent in 200 suspected samples and 40 artificial simulation samples （Kappa=0.927 and 1.000， respectively）.ConclusionThe established RAA-Cas13a detection method has the advantages of high sensitivity and strong specificity. It can quickly detect and screen diarrheal diseases caused by 4 pathogenic bacteria. |
| first_indexed | 2024-03-13T09:46:29Z |
| format | Article |
| id | doaj.art-5f21400f88764fc68e4491f6da9450e0 |
| institution | Directory Open Access Journal |
| issn | 1004-8456 |
| language | zho |
| last_indexed | 2024-03-13T09:46:29Z |
| publishDate | 2023-03-01 |
| publisher | The Editorial Office of Chinese Journal of Food Hygiene |
| record_format | Article |
| series | Zhongguo shipin weisheng zazhi |
| spelling | doaj.art-5f21400f88764fc68e4491f6da9450e02023-05-25T02:13:58ZzhoThe Editorial Office of Chinese Journal of Food HygieneZhongguo shipin weisheng zazhi1004-84562023-03-0135338138910.13590/j.cjfh.2023.03.0101004-8456（2023）03-0381-09Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplificationAN Bailin0SU Xuan1GUO Yue2WANG Xiangxi3GE Yiyue4ZHU Fengcai5CUI Lunbiao6College of Pharmacy, Nankai University, Tianjin 300000, ChinaCollege of Pharmacy, Nankai University, Tianjin 300000, ChinaNanjing Medical University, Jiangsu Nanjing 210029, ChinaCollege of Pharmacy, Nankai University, Tianjin 300000, ChinaNHC Key Laboratory of Enteric Pathogen Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Jiangsu Nanjing 210009, ChinaNHC Key Laboratory of Enteric Pathogen Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Jiangsu Nanjing 210009, ChinaNHC Key Laboratory of Enteric Pathogen Microbiology, Jiangsu Provincial Center for Disease Control and Prevention, Jiangsu Nanjing 210009, ChinaObjectiveTo establish a rapid， sensitive and specific detection method for 4 diarrheal bacteria （Salmonella， Shigella， Vibrio cholera and Escherichia coli O157：H7） by the Clustered regularly interspaced short palindromic repeats associated protein 13a （CRISPR-Cas13a） combined with recombinant enzyme-mediated isothermal amplification （RAA）， called RAA-Cas13a.MethodsIn this study， the specific primer for RAA and CRISPR RNA （crRNA） of 4 different diarrheal bacteria were designed. The sample nucleic acids were amplified by RAA， and the amplification products were then detected with CRISPR-Cas13a. Compared with real-time quantitative polymerase chain reaction（RT-qPCR）， the sensitivity and specificity of the RAA-Cas13a method were evaluated.ResultsThe established RAA-Cas13a detection method for Shigella， Vibrio cholera and Escherichia coli O157：H7 had the detection limit of 10 copies/μL， the detection limit for Salmonella was 1 copy/μL， and each bacteria did not have cross-reaction with the other ten bacteria. Meantime， the detection of the RT-qPCR and RAA-Cas13a were highly consistent in 200 suspected samples and 40 artificial simulation samples （Kappa=0.927 and 1.000， respectively）.ConclusionThe established RAA-Cas13a detection method has the advantages of high sensitivity and strong specificity. It can quickly detect and screen diarrheal diseases caused by 4 pathogenic bacteria.http://www.zgspws.com/zgspwszz/article/abstract/202303010recombinase aided amplificationcrispr-cas13adiarrheal bacteriamolecular detection |
| spellingShingle | AN Bailin SU Xuan GUO Yue WANG Xiangxi GE Yiyue ZHU Fengcai CUI Lunbiao Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplification Zhongguo shipin weisheng zazhi recombinase aided amplification crispr-cas13a diarrheal bacteria molecular detection |
| title | Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplification |
| title_full | Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplification |
| title_fullStr | Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplification |
| title_full_unstemmed | Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplification |
| title_short | Rapid detection of four diarrheal bacteria by CRISPR-Cas13a combined with recombinase aided amplification |
| title_sort | rapid detection of four diarrheal bacteria by crispr cas13a combined with recombinase aided amplification |
| topic | recombinase aided amplification crispr-cas13a diarrheal bacteria molecular detection |
| url | http://www.zgspws.com/zgspwszz/article/abstract/202303010 |
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