A multiplex digital PCR assay for detection and quantitation of porcine circovirus type 2 and type 3
Porcine circovirus (PCV) has become a major pathogen, causing major economic losses in the global pig industry, and PCV type 2 (PCV2) and 3 (PCV3) are distributed worldwide. We designed specific primer and probe sequences targeting PCV2 Cap and PCV3 Rap and developed a multiplex crystal digital PCR...
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| Veröffentlicht in: | Archives of virology 2024-06, Vol.169 (6), p.119-119, Article 119 |
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| creator | Shuai, Jiangbing Chen, Kexin Wang, Zhongcai Zeng, Ruoxue Ma, Biao Zhang, Mingzhou Song, Houhui Zhang, Xiaofeng |
| description | Porcine circovirus (PCV) has become a major pathogen, causing major economic losses in the global pig industry, and PCV type 2 (PCV2) and 3 (PCV3) are distributed worldwide. We designed specific primer and probe sequences targeting PCV2 Cap and PCV3 Rap and developed a multiplex crystal digital PCR (cdPCR) method after optimizing the primer concentration, probe concentration, and annealing temperature. The multiplex cdPCR assay permits precise and differential detection of PCV2 and PCV3, with a limit of detection of 1.39 × 10
1
and 1.27 × 10
1
copies/reaction, respectively, and no cross-reaction with other porcine viruses was observed. The intra-assay and interassay coefficients of variation (CVs) were less than 8.75%, indicating good repeatability and reproducibility. To evaluate the practical value of this assay, 40 tissue samples and 70 feed samples were tested for both PCV2 and PCV3 by cdPCR and quantitative PCR (qPCR). Using multiplex cdPCR, the rates of PCV2 infection, PCV3 infection, and coinfection were 28.45%, 1.72%, and 12.93%, respectively, and using multiplex qPCR, they were 25.00%, 0.86%, and 4.31%, respectively This highly specific and sensitive multiplex cdPCR thus allows accurate simultaneous detection of PCV2 and PCV3, and it is particularly well suited for applications that require the detection of small amounts of input nucleic acid or samples with intensive processing and complex matrices. |
| doi_str_mv | 10.1007/s00705-024-06044-0 |
| format | Article |
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1
and 1.27 × 10
1
copies/reaction, respectively, and no cross-reaction with other porcine viruses was observed. The intra-assay and interassay coefficients of variation (CVs) were less than 8.75%, indicating good repeatability and reproducibility. To evaluate the practical value of this assay, 40 tissue samples and 70 feed samples were tested for both PCV2 and PCV3 by cdPCR and quantitative PCR (qPCR). Using multiplex cdPCR, the rates of PCV2 infection, PCV3 infection, and coinfection were 28.45%, 1.72%, and 12.93%, respectively, and using multiplex qPCR, they were 25.00%, 0.86%, and 4.31%, respectively This highly specific and sensitive multiplex cdPCR thus allows accurate simultaneous detection of PCV2 and PCV3, and it is particularly well suited for applications that require the detection of small amounts of input nucleic acid or samples with intensive processing and complex matrices.</description><identifier>ISSN: 0304-8608</identifier><identifier>EISSN: 1432-8798</identifier><identifier>DOI: 10.1007/s00705-024-06044-0</identifier><identifier>PMID: 38753197</identifier><language>eng</language><publisher>Vienna: Springer Vienna</publisher><subject>Animals ; Biomedical and Life Sciences ; Biomedicine ; Circoviridae Infections - diagnosis ; Circoviridae Infections - veterinary ; Circoviridae Infections - virology ; Circovirus - classification ; Circovirus - genetics ; Circovirus - isolation & purification ; Cross-reaction ; Dermatitis ; DNA Primers - genetics ; DNA, Viral - genetics ; Hogs ; Infections ; Infectious Diseases ; Medical Microbiology ; Multiplex Polymerase Chain Reaction - methods ; Original Article ; Pathogens ; Plasmids ; Polymerase chain reaction ; Quarantine ; Reproducibility of Results ; Reproductive system ; Sensitivity and Specificity ; Swine ; Swine Diseases - diagnosis ; Swine Diseases - virology ; Vaccines ; Virology ; Viruses</subject><ispartof>Archives of virology, 2024-06, Vol.169 (6), p.119-119, Article 119</ispartof><rights>The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.</rights><rights>2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c326t-50aeda468aca4c5b9b48c6100916ca9bac9904cee459a057985deb10b8fd90f93</cites><orcidid>0009-0002-8005-7092</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00705-024-06044-0$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00705-024-06044-0$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,781,785,27928,27929,41492,42561,51323</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/38753197$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Shuai, Jiangbing</creatorcontrib><creatorcontrib>Chen, Kexin</creatorcontrib><creatorcontrib>Wang, Zhongcai</creatorcontrib><creatorcontrib>Zeng, Ruoxue</creatorcontrib><creatorcontrib>Ma, Biao</creatorcontrib><creatorcontrib>Zhang, Mingzhou</creatorcontrib><creatorcontrib>Song, Houhui</creatorcontrib><creatorcontrib>Zhang, Xiaofeng</creatorcontrib><title>A multiplex digital PCR assay for detection and quantitation of porcine circovirus type 2 and type 3</title><title>Archives of virology</title><addtitle>Arch Virol</addtitle><addtitle>Arch Virol</addtitle><description>Porcine circovirus (PCV) has become a major pathogen, causing major economic losses in the global pig industry, and PCV type 2 (PCV2) and 3 (PCV3) are distributed worldwide. We designed specific primer and probe sequences targeting PCV2 Cap and PCV3 Rap and developed a multiplex crystal digital PCR (cdPCR) method after optimizing the primer concentration, probe concentration, and annealing temperature. The multiplex cdPCR assay permits precise and differential detection of PCV2 and PCV3, with a limit of detection of 1.39 × 10
1
and 1.27 × 10
1
copies/reaction, respectively, and no cross-reaction with other porcine viruses was observed. The intra-assay and interassay coefficients of variation (CVs) were less than 8.75%, indicating good repeatability and reproducibility. To evaluate the practical value of this assay, 40 tissue samples and 70 feed samples were tested for both PCV2 and PCV3 by cdPCR and quantitative PCR (qPCR). Using multiplex cdPCR, the rates of PCV2 infection, PCV3 infection, and coinfection were 28.45%, 1.72%, and 12.93%, respectively, and using multiplex qPCR, they were 25.00%, 0.86%, and 4.31%, respectively This highly specific and sensitive multiplex cdPCR thus allows accurate simultaneous detection of PCV2 and PCV3, and it is particularly well suited for applications that require the detection of small amounts of input nucleic acid or samples with intensive processing and complex matrices.</description><subject>Animals</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Circoviridae Infections - diagnosis</subject><subject>Circoviridae Infections - veterinary</subject><subject>Circoviridae Infections - virology</subject><subject>Circovirus - classification</subject><subject>Circovirus - genetics</subject><subject>Circovirus - isolation & purification</subject><subject>Cross-reaction</subject><subject>Dermatitis</subject><subject>DNA Primers - genetics</subject><subject>DNA, Viral - genetics</subject><subject>Hogs</subject><subject>Infections</subject><subject>Infectious Diseases</subject><subject>Medical Microbiology</subject><subject>Multiplex Polymerase Chain Reaction - methods</subject><subject>Original Article</subject><subject>Pathogens</subject><subject>Plasmids</subject><subject>Polymerase chain reaction</subject><subject>Quarantine</subject><subject>Reproducibility of Results</subject><subject>Reproductive system</subject><subject>Sensitivity and Specificity</subject><subject>Swine</subject><subject>Swine Diseases - diagnosis</subject><subject>Swine Diseases - virology</subject><subject>Vaccines</subject><subject>Virology</subject><subject>Viruses</subject><issn>0304-8608</issn><issn>1432-8798</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2024</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU1LxDAQhoMo7vrxBzxIwIuX6rRJ2uS4LH7BgiJ6DmmaLlm6TTdpxf33xq4f4MHLTJh55k0mL0JnKVylAMV1iAFYAhlNIAca4x6appRkCS8E30dTIEATngOfoKMQVgCxQNghmhBeMJKKYoqqGV4PTW-7xrzjyi5trxr8NH_GKgS1xbXzuDK90b11LVZthTeDavtIjQVX4855bVuDtfXavVk_BNxvO4OzkR6P5AQd1KoJ5vQrH6PX25uX-X2yeLx7mM8WiSZZ3icMlKkUzbnSimpWipJyncdVRZprJUqlhQCqjaFMKGBxR1aZMoWS15WAWpBjdLnT7bzbDCb0cm2DNk2jWuOGIAkwxkVWkCKiF3_QlRt8G183UjnjwHmksh2lvQvBm1p23q6V38oU5KcHcueBjB7I0QMJcej8S3oo16b6Gfn-9AiQHRBiq10a_3v3P7IfqfiRWw</recordid><startdate>20240601</startdate><enddate>20240601</enddate><creator>Shuai, Jiangbing</creator><creator>Chen, Kexin</creator><creator>Wang, Zhongcai</creator><creator>Zeng, Ruoxue</creator><creator>Ma, Biao</creator><creator>Zhang, Mingzhou</creator><creator>Song, Houhui</creator><creator>Zhang, Xiaofeng</creator><general>Springer Vienna</general><general>Springer Nature 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>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>K9.</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0009-0002-8005-7092</orcidid></search><sort><creationdate>20240601</creationdate><title>A multiplex digital PCR assay for detection and quantitation of porcine circovirus type 2 and type 3</title><author>Shuai, Jiangbing ; Chen, Kexin ; Wang, Zhongcai ; Zeng, Ruoxue ; Ma, Biao ; Zhang, Mingzhou ; Song, Houhui ; Zhang, Xiaofeng</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c326t-50aeda468aca4c5b9b48c6100916ca9bac9904cee459a057985deb10b8fd90f93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2024</creationdate><topic>Animals</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Circoviridae Infections - diagnosis</topic><topic>Circoviridae Infections - veterinary</topic><topic>Circoviridae Infections - virology</topic><topic>Circovirus - classification</topic><topic>Circovirus - genetics</topic><topic>Circovirus - isolation & purification</topic><topic>Cross-reaction</topic><topic>Dermatitis</topic><topic>DNA Primers - genetics</topic><topic>DNA, Viral - genetics</topic><topic>Hogs</topic><topic>Infections</topic><topic>Infectious Diseases</topic><topic>Medical Microbiology</topic><topic>Multiplex Polymerase Chain Reaction - methods</topic><topic>Original Article</topic><topic>Pathogens</topic><topic>Plasmids</topic><topic>Polymerase chain reaction</topic><topic>Quarantine</topic><topic>Reproducibility of Results</topic><topic>Reproductive system</topic><topic>Sensitivity and Specificity</topic><topic>Swine</topic><topic>Swine Diseases - diagnosis</topic><topic>Swine Diseases - virology</topic><topic>Vaccines</topic><topic>Virology</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Shuai, Jiangbing</creatorcontrib><creatorcontrib>Chen, Kexin</creatorcontrib><creatorcontrib>Wang, Zhongcai</creatorcontrib><creatorcontrib>Zeng, Ruoxue</creatorcontrib><creatorcontrib>Ma, Biao</creatorcontrib><creatorcontrib>Zhang, Mingzhou</creatorcontrib><creatorcontrib>Song, Houhui</creatorcontrib><creatorcontrib>Zhang, Xiaofeng</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Archives of virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Shuai, Jiangbing</au><au>Chen, Kexin</au><au>Wang, Zhongcai</au><au>Zeng, Ruoxue</au><au>Ma, Biao</au><au>Zhang, Mingzhou</au><au>Song, Houhui</au><au>Zhang, Xiaofeng</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A multiplex digital PCR assay for detection and quantitation of porcine circovirus type 2 and type 3</atitle><jtitle>Archives of virology</jtitle><stitle>Arch Virol</stitle><addtitle>Arch Virol</addtitle><date>2024-06-01</date><risdate>2024</risdate><volume>169</volume><issue>6</issue><spage>119</spage><epage>119</epage><pages>119-119</pages><artnum>119</artnum><issn>0304-8608</issn><eissn>1432-8798</eissn><abstract>Porcine circovirus (PCV) has become a major pathogen, causing major economic losses in the global pig industry, and PCV type 2 (PCV2) and 3 (PCV3) are distributed worldwide. We designed specific primer and probe sequences targeting PCV2 Cap and PCV3 Rap and developed a multiplex crystal digital PCR (cdPCR) method after optimizing the primer concentration, probe concentration, and annealing temperature. The multiplex cdPCR assay permits precise and differential detection of PCV2 and PCV3, with a limit of detection of 1.39 × 10
1
and 1.27 × 10
1
copies/reaction, respectively, and no cross-reaction with other porcine viruses was observed. The intra-assay and interassay coefficients of variation (CVs) were less than 8.75%, indicating good repeatability and reproducibility. To evaluate the practical value of this assay, 40 tissue samples and 70 feed samples were tested for both PCV2 and PCV3 by cdPCR and quantitative PCR (qPCR). Using multiplex cdPCR, the rates of PCV2 infection, PCV3 infection, and coinfection were 28.45%, 1.72%, and 12.93%, respectively, and using multiplex qPCR, they were 25.00%, 0.86%, and 4.31%, respectively This highly specific and sensitive multiplex cdPCR thus allows accurate simultaneous detection of PCV2 and PCV3, and it is particularly well suited for applications that require the detection of small amounts of input nucleic acid or samples with intensive processing and complex matrices.</abstract><cop>Vienna</cop><pub>Springer Vienna</pub><pmid>38753197</pmid><doi>10.1007/s00705-024-06044-0</doi><tpages>1</tpages><orcidid>https://orcid.org/0009-0002-8005-7092</orcidid></addata></record> |
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| subjects | Animals Biomedical and Life Sciences Biomedicine Circoviridae Infections - diagnosis Circoviridae Infections - veterinary Circoviridae Infections - virology Circovirus - classification Circovirus - genetics Circovirus - isolation & purification Cross-reaction Dermatitis DNA Primers - genetics DNA, Viral - genetics Hogs Infections Infectious Diseases Medical Microbiology Multiplex Polymerase Chain Reaction - methods Original Article Pathogens Plasmids Polymerase chain reaction Quarantine Reproducibility of Results Reproductive system Sensitivity and Specificity Swine Swine Diseases - diagnosis Swine Diseases - virology Vaccines Virology Viruses |
| title | A multiplex digital PCR assay for detection and quantitation of porcine circovirus type 2 and type 3 |
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