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A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood

Cross-border pathogens such as the African swine fever virus (ASFV) still pose a socio-economic threat. Cheaper, faster, and accurate diagnostics are imperative for healthcare and food safety applications. Currently, the discovery of the Clustered Regularly Interspaced Short Palindromic Repeats (CRI...

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Detalles Bibliográficos
Autores principales: Wu, Jinghua, Mukama, Omar, Wu, Wei, Li, Zhiyuan, Habimana, Jean De Dieu, Zhang, Yinghui, Zeng, Rong, Nie, Chengrong, Zeng, Lingwen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7763806/
https://www.ncbi.nlm.nih.gov/pubmed/33321741
http://dx.doi.org/10.3390/bios10120203
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author Wu, Jinghua
Mukama, Omar
Wu, Wei
Li, Zhiyuan
Habimana, Jean De Dieu
Zhang, Yinghui
Zeng, Rong
Nie, Chengrong
Zeng, Lingwen
author_facet Wu, Jinghua
Mukama, Omar
Wu, Wei
Li, Zhiyuan
Habimana, Jean De Dieu
Zhang, Yinghui
Zeng, Rong
Nie, Chengrong
Zeng, Lingwen
author_sort Wu, Jinghua
collection PubMed
description Cross-border pathogens such as the African swine fever virus (ASFV) still pose a socio-economic threat. Cheaper, faster, and accurate diagnostics are imperative for healthcare and food safety applications. Currently, the discovery of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) has paved the way for the diagnostics based on Cas13 and Cas12/14 that exhibit collateral cleavage of target and single-stranded DNA (ssDNA) reporter. The reporter is fluorescently labeled to report the presence of a target. These methods are powerful; however, fluorescence-based approaches require expensive apparatuses, complicate results readout, and exhibit high-fluorescence background. Here, we present a new CRISPR–Cas-based approach that combines polymerase chain reaction (PCR) amplification, Cas12a, and a probe-based lateral flow biosensor (LFB) for the simultaneous detection of seven types of ASFV. In the presence of ASFVs, the LFB responded to reporter trans-cleavage by naked eyes and achieved a sensitivity of 2.5 × 10(−15) M within 2 h, and unambiguously identified ASFV from swine blood. This system uses less time for PCR pre-amplification and requires cheaper devices; thus, it can be applied to virus monitoring and food samples detection.
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spelling pubmed-77638062020-12-27 A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood Wu, Jinghua Mukama, Omar Wu, Wei Li, Zhiyuan Habimana, Jean De Dieu Zhang, Yinghui Zeng, Rong Nie, Chengrong Zeng, Lingwen Biosensors (Basel) Article Cross-border pathogens such as the African swine fever virus (ASFV) still pose a socio-economic threat. Cheaper, faster, and accurate diagnostics are imperative for healthcare and food safety applications. Currently, the discovery of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) has paved the way for the diagnostics based on Cas13 and Cas12/14 that exhibit collateral cleavage of target and single-stranded DNA (ssDNA) reporter. The reporter is fluorescently labeled to report the presence of a target. These methods are powerful; however, fluorescence-based approaches require expensive apparatuses, complicate results readout, and exhibit high-fluorescence background. Here, we present a new CRISPR–Cas-based approach that combines polymerase chain reaction (PCR) amplification, Cas12a, and a probe-based lateral flow biosensor (LFB) for the simultaneous detection of seven types of ASFV. In the presence of ASFVs, the LFB responded to reporter trans-cleavage by naked eyes and achieved a sensitivity of 2.5 × 10(−15) M within 2 h, and unambiguously identified ASFV from swine blood. This system uses less time for PCR pre-amplification and requires cheaper devices; thus, it can be applied to virus monitoring and food samples detection. MDPI 2020-12-10 /pmc/articles/PMC7763806/ /pubmed/33321741 http://dx.doi.org/10.3390/bios10120203 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Wu, Jinghua
Mukama, Omar
Wu, Wei
Li, Zhiyuan
Habimana, Jean De Dieu
Zhang, Yinghui
Zeng, Rong
Nie, Chengrong
Zeng, Lingwen
A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood
title A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood
title_full A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood
title_fullStr A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood
title_full_unstemmed A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood
title_short A CRISPR/Cas12a Based Universal Lateral Flow Biosensor for the Sensitive and Specific Detection of African Swine-Fever Viruses in Whole Blood
title_sort crispr/cas12a based universal lateral flow biosensor for the sensitive and specific detection of african swine-fever viruses in whole blood
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7763806/
https://www.ncbi.nlm.nih.gov/pubmed/33321741
http://dx.doi.org/10.3390/bios10120203
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