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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...

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Autores principales: Huang, Mengqian, Liu, Sihua, Xu, Yanan, Li, Aqian, Wu, Wei, Liang, Mifang, Niu, Guoyu, Wang, Zhiyun, Wang, Tao
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
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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.
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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
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