Cargando…
Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay
BACKGROUND: Integrating CRISPR-Cas12a sensors with isothermal signal amplification can be exploited to develop low-cost, disposable, and ultrasensitive assays for the diagnostics of human pathogens. METHODS: RT-RAA-Cas12a-mediated real-time or end-point fluorescent and lateral flow strip (LFS) assay...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9482751/ https://www.ncbi.nlm.nih.gov/pubmed/36115975 http://dx.doi.org/10.1186/s12985-022-01878-z |
_version_ | 1784791523617931264 |
---|---|
author | Qian, Weidong Huang, Jie Wang, Ting Fan, Cheng Kang, Jie Zhang, Qian Li, Yongdong Chen, Si |
author_facet | Qian, Weidong Huang, Jie Wang, Ting Fan, Cheng Kang, Jie Zhang, Qian Li, Yongdong Chen, Si |
author_sort | Qian, Weidong |
collection | PubMed |
description | BACKGROUND: Integrating CRISPR-Cas12a sensors with isothermal signal amplification can be exploited to develop low-cost, disposable, and ultrasensitive assays for the diagnostics of human pathogens. METHODS: RT-RAA-Cas12a-mediated real-time or end-point fluorescent and lateral flow strip (LFS) assays for direct detection of norovirus (NOV) genotype GII.4 or GII.17 were explored. RESULTS: The results showed that our RT-RAA-Cas12a-mediated fluorescent and LFS assay could detect NOV GII.4 or GII.17 by targeting the viral protein 1 gene. Our RT-RAA-Cas12a-mediated fluorescent and LFS assay can specifically detect NOV GII.4 or GII.17 with no cross-reactivity for other related viruses. The low limit of detection could reach 0.1 copies/μL within approximately 30–40 min, and the results were visualized using an ultraviolet light illuminator or on a LFS without complex equipment. In addition, our RT-RAA-Cas12a-mediated fluorescent and LFS assay provided a visual and faster alternative to real-time RT-PCR assay, with 95.7% and 94.3% positive predictive agreement and 100% negative predictive agreement. CONCLUSIONS: Together, our RT-RAA-Cas12a-mediated approach would have a great potential for point-of-care diagnostics of NOV GII.4 and/or GII.17 in resource-limited settings. |
format | Online Article Text |
id | pubmed-9482751 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-94827512022-09-19 Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay Qian, Weidong Huang, Jie Wang, Ting Fan, Cheng Kang, Jie Zhang, Qian Li, Yongdong Chen, Si Virol J Research BACKGROUND: Integrating CRISPR-Cas12a sensors with isothermal signal amplification can be exploited to develop low-cost, disposable, and ultrasensitive assays for the diagnostics of human pathogens. METHODS: RT-RAA-Cas12a-mediated real-time or end-point fluorescent and lateral flow strip (LFS) assays for direct detection of norovirus (NOV) genotype GII.4 or GII.17 were explored. RESULTS: The results showed that our RT-RAA-Cas12a-mediated fluorescent and LFS assay could detect NOV GII.4 or GII.17 by targeting the viral protein 1 gene. Our RT-RAA-Cas12a-mediated fluorescent and LFS assay can specifically detect NOV GII.4 or GII.17 with no cross-reactivity for other related viruses. The low limit of detection could reach 0.1 copies/μL within approximately 30–40 min, and the results were visualized using an ultraviolet light illuminator or on a LFS without complex equipment. In addition, our RT-RAA-Cas12a-mediated fluorescent and LFS assay provided a visual and faster alternative to real-time RT-PCR assay, with 95.7% and 94.3% positive predictive agreement and 100% negative predictive agreement. CONCLUSIONS: Together, our RT-RAA-Cas12a-mediated approach would have a great potential for point-of-care diagnostics of NOV GII.4 and/or GII.17 in resource-limited settings. BioMed Central 2022-09-17 /pmc/articles/PMC9482751/ /pubmed/36115975 http://dx.doi.org/10.1186/s12985-022-01878-z Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Qian, Weidong Huang, Jie Wang, Ting Fan, Cheng Kang, Jie Zhang, Qian Li, Yongdong Chen, Si Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay |
title | Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay |
title_full | Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay |
title_fullStr | Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay |
title_full_unstemmed | Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay |
title_short | Ultrasensitive and visual detection of human norovirus genotype GII.4 or GII.17 using CRISPR-Cas12a assay |
title_sort | ultrasensitive and visual detection of human norovirus genotype gii.4 or gii.17 using crispr-cas12a assay |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9482751/ https://www.ncbi.nlm.nih.gov/pubmed/36115975 http://dx.doi.org/10.1186/s12985-022-01878-z |
work_keys_str_mv | AT qianweidong ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay AT huangjie ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay AT wangting ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay AT fancheng ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay AT kangjie ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay AT zhangqian ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay AT liyongdong ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay AT chensi ultrasensitiveandvisualdetectionofhumannorovirusgenotypegii4orgii17usingcrisprcas12aassay |