Development of visual detection of African swine fever virus using CRISPR/LwCas13a lateral flow strip based on structural protein gene D117L
African swine fever virus (ASFV) is a large double stranded DNA arbovirus that is highly contagious and seriously endangers domestic and wild pigs. In the past decade, African swine fever (ASF) has spread in many countries in the Caucasus, Russian Federation, Eastern Europe and Asia, causing signifi...
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creator | Zhang, Desheng Jiang, Sen Xia, Nengwen Zhang, Jiajia Liu, Anjing Deng, Dafu Zhang, Chenyang Sun, Yuxin Chen, Nanhua Kang, Xilong Pan, Zhiming Zheng, Wanglong Zhu, Jianzhong |
description | African swine fever virus (ASFV) is a large double stranded DNA arbovirus that is highly contagious and seriously endangers domestic and wild pigs. In the past decade, African swine fever (ASF) has spread in many countries in the Caucasus, Russian Federation, Eastern Europe and Asia, causing significant losses to the pig industry. At present, there is a lack of effective vaccine and treatment for ASF. Therefore, the rapid and accurate detection is crucial for ASF prevention and control. In this study, we have developed a portable lateral flow strip (LFS) detection mediated by recombinase polymerase amplification (RPA) and CRISPR/LwCas13a, which is performed at 37 ℃ and visualized by eyes without the need for complex instruments. This RPA-LwCas13a-LFS is based on the ASFV structural protein p17 gene (D117L), with a detection sensitivity up to 2 gene copies. This method is highly specific and has no cross reactivity to 7 other pig viruses. In the detection of two batches of 100 clinical samples, the p17 (D117L) RPA-LwCas13a-LFS had 100% coincidence with conventional quantitative PCR (qPCR). These findings demonstrate the potential of this simple, rapid, sensitive, and specific ASFV detection method for on-site ASFV detection.
•The RPA-Cas13a-LFS detection method was developed based on ASFV p17 gene.•The LFS method exhibited a high sensitivity and specificity in detection of ASFV.•The RPA-LwCas13a-LFS had 100% coincidence with conventional qPCR in the detection of 100 clinical samples. |
doi_str_mv | 10.1016/j.vetmic.2024.110073 |
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•The RPA-Cas13a-LFS detection method was developed based on ASFV p17 gene.•The LFS method exhibited a high sensitivity and specificity in detection of ASFV.•The RPA-LwCas13a-LFS had 100% coincidence with conventional qPCR in the detection of 100 clinical samples.</description><identifier>ISSN: 0378-1135</identifier><identifier>ISSN: 1873-2542</identifier><identifier>EISSN: 1873-2542</identifier><identifier>DOI: 10.1016/j.vetmic.2024.110073</identifier><identifier>PMID: 38579481</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>African Swine Fever - diagnosis ; African Swine Fever - virology ; African swine fever virus (ASFV) ; African Swine Fever Virus - genetics ; African Swine Fever Virus - isolation & purification ; Animals ; CRISPR-Cas Systems ; CRISPR/LwCas13a ; Lateral flow strip (LFS) ; Nucleic Acid Amplification Techniques - methods ; Nucleic Acid Amplification Techniques - veterinary ; Recombinase polymerase amplification (RPA) ; Sensitivity and Specificity ; Swine ; Viral Structural Proteins - genetics ; Visual detection</subject><ispartof>Veterinary microbiology, 2024-06, Vol.293, p.110073, Article 110073</ispartof><rights>2024 Elsevier B.V.</rights><rights>Copyright © 2024 Elsevier B.V. All rights reserved.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c357t-425765ccc75f40893879ae1ae42e6bde7ca193d8b6953fc98bcacbee7f4301e43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.vetmic.2024.110073$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38579481$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Desheng</creatorcontrib><creatorcontrib>Jiang, Sen</creatorcontrib><creatorcontrib>Xia, Nengwen</creatorcontrib><creatorcontrib>Zhang, Jiajia</creatorcontrib><creatorcontrib>Liu, Anjing</creatorcontrib><creatorcontrib>Deng, Dafu</creatorcontrib><creatorcontrib>Zhang, Chenyang</creatorcontrib><creatorcontrib>Sun, Yuxin</creatorcontrib><creatorcontrib>Chen, Nanhua</creatorcontrib><creatorcontrib>Kang, Xilong</creatorcontrib><creatorcontrib>Pan, Zhiming</creatorcontrib><creatorcontrib>Zheng, Wanglong</creatorcontrib><creatorcontrib>Zhu, Jianzhong</creatorcontrib><title>Development of visual detection of African swine fever virus using CRISPR/LwCas13a lateral flow strip based on structural protein gene D117L</title><title>Veterinary microbiology</title><addtitle>Vet Microbiol</addtitle><description>African swine fever virus (ASFV) is a large double stranded DNA arbovirus that is highly contagious and seriously endangers domestic and wild pigs. In the past decade, African swine fever (ASF) has spread in many countries in the Caucasus, Russian Federation, Eastern Europe and Asia, causing significant losses to the pig industry. At present, there is a lack of effective vaccine and treatment for ASF. Therefore, the rapid and accurate detection is crucial for ASF prevention and control. In this study, we have developed a portable lateral flow strip (LFS) detection mediated by recombinase polymerase amplification (RPA) and CRISPR/LwCas13a, which is performed at 37 ℃ and visualized by eyes without the need for complex instruments. This RPA-LwCas13a-LFS is based on the ASFV structural protein p17 gene (D117L), with a detection sensitivity up to 2 gene copies. This method is highly specific and has no cross reactivity to 7 other pig viruses. In the detection of two batches of 100 clinical samples, the p17 (D117L) RPA-LwCas13a-LFS had 100% coincidence with conventional quantitative PCR (qPCR). These findings demonstrate the potential of this simple, rapid, sensitive, and specific ASFV detection method for on-site ASFV detection.
•The RPA-Cas13a-LFS detection method was developed based on ASFV p17 gene.•The LFS method exhibited a high sensitivity and specificity in detection of ASFV.•The RPA-LwCas13a-LFS had 100% coincidence with conventional qPCR in the detection of 100 clinical samples.</description><subject>African Swine Fever - diagnosis</subject><subject>African Swine Fever - virology</subject><subject>African swine fever virus (ASFV)</subject><subject>African Swine Fever Virus - genetics</subject><subject>African Swine Fever Virus - isolation & purification</subject><subject>Animals</subject><subject>CRISPR-Cas Systems</subject><subject>CRISPR/LwCas13a</subject><subject>Lateral flow strip (LFS)</subject><subject>Nucleic Acid Amplification Techniques - methods</subject><subject>Nucleic Acid Amplification Techniques - veterinary</subject><subject>Recombinase polymerase amplification (RPA)</subject><subject>Sensitivity and Specificity</subject><subject>Swine</subject><subject>Viral Structural Proteins - genetics</subject><subject>Visual detection</subject><issn>0378-1135</issn><issn>1873-2542</issn><issn>1873-2542</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1uGyEUhVHVqHHcvEFVsexmHBhgmNlUiuw2sWQpUdKsEcNcLKz5cYGxlXfIQwdrkiyzQhe-cy73HoR-ULKghBZXu8UBYufMIic5X1BKiGRf0IyWkmW54PlXNCNMlhmlTJyjixB2hBBeFeQbOmelkBUv6Qy9rOAA7bDvoI94sPjgwqhb3EAEE93Qn-6urXdG9zgcXQ_YJoFPnB8DHoPrt3j5sH68f7jaHJc6UKZxqyP4ZGLb4YhD9G6Pax2gwckulaOJ4-l574cIrsdbSK4rSuXmOzqzug1w-XbO0dPfP_-Wt9nm7ma9vN5khgkZM54LWQhjjBSWk7Jipaw0UA08h6JuQBpNK9aUdVEJZk1V1kabGkBazggFzubo1-SbvvB_hBBV54KBttU9DGNQjDCe84qLIqF8Qo0fQvBg1d67TvtnRYk65aB2aspBnXJQUw5J9vOtw1h30HyI3hefgN8TAGnOgwOvgnHQG2icT5tXzeA-7_AKhjycjg</recordid><startdate>202406</startdate><enddate>202406</enddate><creator>Zhang, Desheng</creator><creator>Jiang, Sen</creator><creator>Xia, Nengwen</creator><creator>Zhang, Jiajia</creator><creator>Liu, Anjing</creator><creator>Deng, Dafu</creator><creator>Zhang, Chenyang</creator><creator>Sun, Yuxin</creator><creator>Chen, Nanhua</creator><creator>Kang, Xilong</creator><creator>Pan, Zhiming</creator><creator>Zheng, Wanglong</creator><creator>Zhu, Jianzhong</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>202406</creationdate><title>Development of visual detection of African swine fever virus using CRISPR/LwCas13a lateral flow strip based on structural protein gene D117L</title><author>Zhang, Desheng ; Jiang, Sen ; Xia, Nengwen ; Zhang, Jiajia ; Liu, Anjing ; Deng, Dafu ; Zhang, Chenyang ; Sun, Yuxin ; Chen, Nanhua ; Kang, Xilong ; Pan, Zhiming ; Zheng, Wanglong ; Zhu, Jianzhong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c357t-425765ccc75f40893879ae1ae42e6bde7ca193d8b6953fc98bcacbee7f4301e43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>African Swine Fever - diagnosis</topic><topic>African Swine Fever - virology</topic><topic>African swine fever virus (ASFV)</topic><topic>African Swine Fever Virus - genetics</topic><topic>African Swine Fever Virus - isolation & purification</topic><topic>Animals</topic><topic>CRISPR-Cas Systems</topic><topic>CRISPR/LwCas13a</topic><topic>Lateral flow strip (LFS)</topic><topic>Nucleic Acid Amplification Techniques - methods</topic><topic>Nucleic Acid Amplification Techniques - veterinary</topic><topic>Recombinase polymerase amplification (RPA)</topic><topic>Sensitivity and Specificity</topic><topic>Swine</topic><topic>Viral Structural Proteins - genetics</topic><topic>Visual detection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Desheng</creatorcontrib><creatorcontrib>Jiang, Sen</creatorcontrib><creatorcontrib>Xia, Nengwen</creatorcontrib><creatorcontrib>Zhang, Jiajia</creatorcontrib><creatorcontrib>Liu, Anjing</creatorcontrib><creatorcontrib>Deng, Dafu</creatorcontrib><creatorcontrib>Zhang, Chenyang</creatorcontrib><creatorcontrib>Sun, Yuxin</creatorcontrib><creatorcontrib>Chen, Nanhua</creatorcontrib><creatorcontrib>Kang, Xilong</creatorcontrib><creatorcontrib>Pan, Zhiming</creatorcontrib><creatorcontrib>Zheng, Wanglong</creatorcontrib><creatorcontrib>Zhu, Jianzhong</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Veterinary microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Desheng</au><au>Jiang, Sen</au><au>Xia, Nengwen</au><au>Zhang, Jiajia</au><au>Liu, Anjing</au><au>Deng, Dafu</au><au>Zhang, Chenyang</au><au>Sun, Yuxin</au><au>Chen, Nanhua</au><au>Kang, Xilong</au><au>Pan, Zhiming</au><au>Zheng, Wanglong</au><au>Zhu, Jianzhong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of visual detection of African swine fever virus using CRISPR/LwCas13a lateral flow strip based on structural protein gene D117L</atitle><jtitle>Veterinary microbiology</jtitle><addtitle>Vet Microbiol</addtitle><date>2024-06</date><risdate>2024</risdate><volume>293</volume><spage>110073</spage><pages>110073-</pages><artnum>110073</artnum><issn>0378-1135</issn><issn>1873-2542</issn><eissn>1873-2542</eissn><abstract>African swine fever virus (ASFV) is a large double stranded DNA arbovirus that is highly contagious and seriously endangers domestic and wild pigs. In the past decade, African swine fever (ASF) has spread in many countries in the Caucasus, Russian Federation, Eastern Europe and Asia, causing significant losses to the pig industry. At present, there is a lack of effective vaccine and treatment for ASF. Therefore, the rapid and accurate detection is crucial for ASF prevention and control. In this study, we have developed a portable lateral flow strip (LFS) detection mediated by recombinase polymerase amplification (RPA) and CRISPR/LwCas13a, which is performed at 37 ℃ and visualized by eyes without the need for complex instruments. This RPA-LwCas13a-LFS is based on the ASFV structural protein p17 gene (D117L), with a detection sensitivity up to 2 gene copies. This method is highly specific and has no cross reactivity to 7 other pig viruses. In the detection of two batches of 100 clinical samples, the p17 (D117L) RPA-LwCas13a-LFS had 100% coincidence with conventional quantitative PCR (qPCR). These findings demonstrate the potential of this simple, rapid, sensitive, and specific ASFV detection method for on-site ASFV detection.
•The RPA-Cas13a-LFS detection method was developed based on ASFV p17 gene.•The LFS method exhibited a high sensitivity and specificity in detection of ASFV.•The RPA-LwCas13a-LFS had 100% coincidence with conventional qPCR in the detection of 100 clinical samples.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>38579481</pmid><doi>10.1016/j.vetmic.2024.110073</doi><oa>free_for_read</oa></addata></record> |
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subjects | African Swine Fever - diagnosis African Swine Fever - virology African swine fever virus (ASFV) African Swine Fever Virus - genetics African Swine Fever Virus - isolation & purification Animals CRISPR-Cas Systems CRISPR/LwCas13a Lateral flow strip (LFS) Nucleic Acid Amplification Techniques - methods Nucleic Acid Amplification Techniques - veterinary Recombinase polymerase amplification (RPA) Sensitivity and Specificity Swine Viral Structural Proteins - genetics Visual detection |
title | Development of visual detection of African swine fever virus using CRISPR/LwCas13a lateral flow strip based on structural protein gene D117L |
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