Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Lu, Pan, Chen, Jialiang, Li, Zhenpeng, Li, Zhe, Zhang, Jingyun, Kan, Biao, Pang, Bo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
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
_version_ 1784687085932773376
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
work_keys_str_mv AT lupan visualidentificationandserotypingoftoxigenicvibriocholeraeserogroupso1ando139withcarid
AT chenjialiang visualidentificationandserotypingoftoxigenicvibriocholeraeserogroupso1ando139withcarid
AT lizhenpeng visualidentificationandserotypingoftoxigenicvibriocholeraeserogroupso1ando139withcarid
AT lizhe visualidentificationandserotypingoftoxigenicvibriocholeraeserogroupso1ando139withcarid
AT zhangjingyun visualidentificationandserotypingoftoxigenicvibriocholeraeserogroupso1ando139withcarid
AT kanbiao visualidentificationandserotypingoftoxigenicvibriocholeraeserogroupso1ando139withcarid
AT pangbo visualidentificationandserotypingoftoxigenicvibriocholeraeserogroupso1ando139withcarid