Cargando…
CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection
Severe fever with thrombocytopenia syndrome virus (SFTSV) is a new tick-borne pathogen that can cause severe hemorrhagic fever. Fever with thrombocytopenia syndrome caused by SFTSV is a new infectious disease that has posed a great threat to public health. Therefore, a fast, sensitive, low-cost, and...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8822122/ https://www.ncbi.nlm.nih.gov/pubmed/35145502 http://dx.doi.org/10.3389/fmicb.2022.754995 |
_version_ | 1784646546896191488 |
---|---|
author | Huang, Mengqian Liu, Sihua Xu, Yanan Li, Aqian Wu, Wei Liang, Mifang Niu, Guoyu Wang, Zhiyun Wang, Tao |
author_facet | Huang, Mengqian Liu, Sihua Xu, Yanan Li, Aqian Wu, Wei Liang, Mifang Niu, Guoyu Wang, Zhiyun Wang, Tao |
author_sort | Huang, Mengqian |
collection | PubMed |
description | Severe fever with thrombocytopenia syndrome virus (SFTSV) is a new tick-borne pathogen that can cause severe hemorrhagic fever. Fever with thrombocytopenia syndrome caused by SFTSV is a new infectious disease that has posed a great threat to public health. Therefore, a fast, sensitive, low-cost, and field-deployable detection method for diagnosing SFTSV is essential for virus surveillance and control. In this study, we developed a rapid, highly sensitive, instrument-flexible SFTSV detection method that utilizes recombinase polymerase amplification and the CRISPR/Cas12a system. We found that three copies of the L gene from the SFTSV genome per reaction were enough to ensure stable detection within 40 min. The assay clearly showed no cross-reactivity with other RNA viruses. Additionally, our method demonstrated 100% agreement with Q-PCR detection results for SFTSV in 46 clinical samples. We simplified the requirements for on-site detection instruments by combining the CRISPR/Cas12a tool and immunochromatographic strips to create a system that can reliably detect one copy/μl sample of the L gene, which showed extremely high sensitivity and specificity for detecting the virus. Taken together, these findings indicate that the new SFTSV detection method is a powerful and effective tool for on-site detection, which can contribute to diagnosing SFTSV quickly and sensitively. |
format | Online Article Text |
id | pubmed-8822122 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-88221222022-02-09 CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection Huang, Mengqian Liu, Sihua Xu, Yanan Li, Aqian Wu, Wei Liang, Mifang Niu, Guoyu Wang, Zhiyun Wang, Tao Front Microbiol Microbiology Severe fever with thrombocytopenia syndrome virus (SFTSV) is a new tick-borne pathogen that can cause severe hemorrhagic fever. Fever with thrombocytopenia syndrome caused by SFTSV is a new infectious disease that has posed a great threat to public health. Therefore, a fast, sensitive, low-cost, and field-deployable detection method for diagnosing SFTSV is essential for virus surveillance and control. In this study, we developed a rapid, highly sensitive, instrument-flexible SFTSV detection method that utilizes recombinase polymerase amplification and the CRISPR/Cas12a system. We found that three copies of the L gene from the SFTSV genome per reaction were enough to ensure stable detection within 40 min. The assay clearly showed no cross-reactivity with other RNA viruses. Additionally, our method demonstrated 100% agreement with Q-PCR detection results for SFTSV in 46 clinical samples. We simplified the requirements for on-site detection instruments by combining the CRISPR/Cas12a tool and immunochromatographic strips to create a system that can reliably detect one copy/μl sample of the L gene, which showed extremely high sensitivity and specificity for detecting the virus. Taken together, these findings indicate that the new SFTSV detection method is a powerful and effective tool for on-site detection, which can contribute to diagnosing SFTSV quickly and sensitively. Frontiers Media S.A. 2022-01-25 /pmc/articles/PMC8822122/ /pubmed/35145502 http://dx.doi.org/10.3389/fmicb.2022.754995 Text en Copyright © 2022 Huang, Liu, Xu, Li, Wu, Liang, Niu, Wang and Wang. 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 | Microbiology Huang, Mengqian Liu, Sihua Xu, Yanan Li, Aqian Wu, Wei Liang, Mifang Niu, Guoyu Wang, Zhiyun Wang, Tao CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection |
title | CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection |
title_full | CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection |
title_fullStr | CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection |
title_full_unstemmed | CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection |
title_short | CRISPR/Cas12a Technology Combined With RPA for Rapid and Portable SFTSV Detection |
title_sort | crispr/cas12a technology combined with rpa for rapid and portable sftsv detection |
topic | Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8822122/ https://www.ncbi.nlm.nih.gov/pubmed/35145502 http://dx.doi.org/10.3389/fmicb.2022.754995 |
work_keys_str_mv | AT huangmengqian crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT liusihua crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT xuyanan crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT liaqian crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT wuwei crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT liangmifang crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT niuguoyu crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT wangzhiyun crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection AT wangtao crisprcas12atechnologycombinedwithrpaforrapidandportablesftsvdetection |