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Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID
There is a growing demand for rapid, sensitive, field-deployable nucleic acid tests for cholera, which usually occurs in rural areas. In this study, we developed a Cas12a-assisted rapid isothermal detection (CARID) system for the detection of toxigenic V. cholerae serogroups O1 and O139 by combining...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9008587/ https://www.ncbi.nlm.nih.gov/pubmed/35433512 http://dx.doi.org/10.3389/fcimb.2022.863435 |
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author | Lu, Pan Chen, Jialiang Li, Zhenpeng Li, Zhe Zhang, Jingyun Kan, Biao Pang, Bo |
author_facet | Lu, Pan Chen, Jialiang Li, Zhenpeng Li, Zhe Zhang, Jingyun Kan, Biao Pang, Bo |
author_sort | Lu, Pan |
collection | PubMed |
description | There is a growing demand for rapid, sensitive, field-deployable nucleic acid tests for cholera, which usually occurs in rural areas. In this study, we developed a Cas12a-assisted rapid isothermal detection (CARID) system for the detection of toxigenic V. cholerae serogroups O1 and O139 by combining recombinase-aided amplification and CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins). The results can be determined by fluorescence signal and visualized by lateral flow dipstick. We identified 154 V. cholerae strains and 129 strains of other intestinal diarrheagenic bacteria with a 100% coincidence rate. The limit of detection of CARID was 20 copies/reaction of V. cholerae genomic DNA, which is comparable to that of polymerase chain reaction (PCR) and qPCR. Multiple-CARID was also established for efficiency and economic considerations with an acceptable decrease in sensitivity. Simulated sample tests showed that CARID is suitable for complex samples. In conclusion, CARID is a rapid, sensitive, economically efficient, and portable method for the detection of V. cholerae, which makes it suitable for field responses to cholera. |
format | Online Article Text |
id | pubmed-9008587 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-90085872022-04-15 Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID Lu, Pan Chen, Jialiang Li, Zhenpeng Li, Zhe Zhang, Jingyun Kan, Biao Pang, Bo Front Cell Infect Microbiol Cellular and Infection Microbiology There is a growing demand for rapid, sensitive, field-deployable nucleic acid tests for cholera, which usually occurs in rural areas. In this study, we developed a Cas12a-assisted rapid isothermal detection (CARID) system for the detection of toxigenic V. cholerae serogroups O1 and O139 by combining recombinase-aided amplification and CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins). The results can be determined by fluorescence signal and visualized by lateral flow dipstick. We identified 154 V. cholerae strains and 129 strains of other intestinal diarrheagenic bacteria with a 100% coincidence rate. The limit of detection of CARID was 20 copies/reaction of V. cholerae genomic DNA, which is comparable to that of polymerase chain reaction (PCR) and qPCR. Multiple-CARID was also established for efficiency and economic considerations with an acceptable decrease in sensitivity. Simulated sample tests showed that CARID is suitable for complex samples. In conclusion, CARID is a rapid, sensitive, economically efficient, and portable method for the detection of V. cholerae, which makes it suitable for field responses to cholera. Frontiers Media S.A. 2022-03-31 /pmc/articles/PMC9008587/ /pubmed/35433512 http://dx.doi.org/10.3389/fcimb.2022.863435 Text en Copyright © 2022 Lu, Chen, Li, Li, Zhang, Kan and Pang https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cellular and Infection Microbiology Lu, Pan Chen, Jialiang Li, Zhenpeng Li, Zhe Zhang, Jingyun Kan, Biao Pang, Bo Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID |
title | Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID |
title_full | Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID |
title_fullStr | Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID |
title_full_unstemmed | Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID |
title_short | Visual Identification and Serotyping of Toxigenic Vibrio cholerae Serogroups O1 and O139 With CARID |
title_sort | visual identification and serotyping of toxigenic vibrio cholerae serogroups o1 and o139 with carid |
topic | Cellular and Infection Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9008587/ https://www.ncbi.nlm.nih.gov/pubmed/35433512 http://dx.doi.org/10.3389/fcimb.2022.863435 |
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