Establishment of TaqMan-based real-time PCR assay for rapid detection of duck circovirus
Duck circovirus (DuCV) is widespread across the world and causes feather disorders in young ducks. It was identified as the causative pathogen of duck beak atrophy and dwarfism syndrome and primary sclerosing cholangitis. In this study, we aimed to establish a TaqMan-based real-time PCR assay to det...
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description | Duck circovirus (DuCV) is widespread across the world and causes feather disorders in young ducks. It was identified as the causative pathogen of duck beak atrophy and dwarfism syndrome and primary sclerosing cholangitis. In this study, we aimed to establish a TaqMan-based real-time PCR assay to detect DuCV. The primers and probe were designed based on the conserved region of the DuCV Rep gene. After optimizing the reaction conditions, the minimum virus detection limit of the designed PCR technique was 39.4 copies/μL, 100 times that of conventional PCR (cPCR). No cross-reaction with six other common duck viruses was observed. The intra- and inter-assay variations were less than 1%. The detection rate of DuCV-positive clinical samples using TaqMan-based real-time PCR was higher than that using SYBR Green-based real-time PCR and cPCR. Collectively, these results showed that the established TaqMan-based real-time PCR detected DuCV with high sensitivity and specificity, and significant repeatability, making it suitable for clinical use. Hence, it may be used as a novel tool for the diagnosis and epidemiological investigation of DuCV. |
doi_str_mv | 10.1007/s13205-021-03021-1 |
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It was identified as the causative pathogen of duck beak atrophy and dwarfism syndrome and primary sclerosing cholangitis. In this study, we aimed to establish a TaqMan-based real-time PCR assay to detect DuCV. The primers and probe were designed based on the conserved region of the DuCV Rep gene. After optimizing the reaction conditions, the minimum virus detection limit of the designed PCR technique was 39.4 copies/μL, 100 times that of conventional PCR (cPCR). No cross-reaction with six other common duck viruses was observed. The intra- and inter-assay variations were less than 1%. The detection rate of DuCV-positive clinical samples using TaqMan-based real-time PCR was higher than that using SYBR Green-based real-time PCR and cPCR. Collectively, these results showed that the established TaqMan-based real-time PCR detected DuCV with high sensitivity and specificity, and significant repeatability, making it suitable for clinical use. Hence, it may be used as a novel tool for the diagnosis and epidemiological investigation of DuCV.</description><identifier>ISSN: 2190-572X</identifier><identifier>EISSN: 2190-5738</identifier><identifier>DOI: 10.1007/s13205-021-03021-1</identifier><identifier>PMID: 34745821</identifier><language>eng</language><publisher>Cham: Springer International Publishing</publisher><subject>Agriculture ; Assaying ; Atrophy ; Bioinformatics ; Biomaterials ; Biotechnology ; Cancer Research ; Chemistry ; Chemistry and Materials Science ; Cholangitis ; Cross-reaction ; Dwarfism ; Epidemiology ; Original ; Original Article ; Real time ; Rep gene ; Stem Cells ; Waterfowl</subject><ispartof>3 Biotech, 2021-11, Vol.11 (11), p.470-470, Article 470</ispartof><rights>King Abdulaziz City for Science and Technology 2021</rights><rights>King Abdulaziz City for Science and Technology 2021.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c451t-becf63e80cf91c5a5c4bb95eda38cb8d80ac7dc2ebf9e74ef6c5c67ac8b85eb13</citedby><cites>FETCH-LOGICAL-c451t-becf63e80cf91c5a5c4bb95eda38cb8d80ac7dc2ebf9e74ef6c5c67ac8b85eb13</cites><orcidid>0000-0001-8724-0370</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8536814/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8536814/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,41488,42557,51319,53791,53793</link.rule.ids></links><search><creatorcontrib>Zhang, Da</creatorcontrib><creatorcontrib>Wu, Junhuang</creatorcontrib><creatorcontrib>Sun, Jianfei</creatorcontrib><creatorcontrib>Bai, Caixia</creatorcontrib><creatorcontrib>Xu, Fazhi</creatorcontrib><creatorcontrib>Duan, Pengfei</creatorcontrib><creatorcontrib>Wang, Yong</creatorcontrib><title>Establishment of TaqMan-based real-time PCR assay for rapid detection of duck circovirus</title><title>3 Biotech</title><addtitle>3 Biotech</addtitle><description>Duck circovirus (DuCV) is widespread across the world and causes feather disorders in young ducks. It was identified as the causative pathogen of duck beak atrophy and dwarfism syndrome and primary sclerosing cholangitis. In this study, we aimed to establish a TaqMan-based real-time PCR assay to detect DuCV. The primers and probe were designed based on the conserved region of the DuCV Rep gene. After optimizing the reaction conditions, the minimum virus detection limit of the designed PCR technique was 39.4 copies/μL, 100 times that of conventional PCR (cPCR). No cross-reaction with six other common duck viruses was observed. The intra- and inter-assay variations were less than 1%. The detection rate of DuCV-positive clinical samples using TaqMan-based real-time PCR was higher than that using SYBR Green-based real-time PCR and cPCR. Collectively, these results showed that the established TaqMan-based real-time PCR detected DuCV with high sensitivity and specificity, and significant repeatability, making it suitable for clinical use. Hence, it may be used as a novel tool for the diagnosis and epidemiological investigation of DuCV.</description><subject>Agriculture</subject><subject>Assaying</subject><subject>Atrophy</subject><subject>Bioinformatics</subject><subject>Biomaterials</subject><subject>Biotechnology</subject><subject>Cancer Research</subject><subject>Chemistry</subject><subject>Chemistry and Materials Science</subject><subject>Cholangitis</subject><subject>Cross-reaction</subject><subject>Dwarfism</subject><subject>Epidemiology</subject><subject>Original</subject><subject>Original Article</subject><subject>Real time</subject><subject>Rep gene</subject><subject>Stem Cells</subject><subject>Waterfowl</subject><issn>2190-572X</issn><issn>2190-5738</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><recordid>eNp9kU9L5TAUxcMwwyjqF5hVYDazqZPbNG26EeTh_AFFGRxwF5LbW43TNs-kFfz25s0TRRdmkQTyO4ecexj7AuIQhGi-J5ClUIUooRBys8MHtltCKwrVSP3x-V5e7bCDlG5FXgpUC-Iz25FVUyldwi67OkmzdYNPNyNNMw89v7R3Z3YqnE3U8Uh2KGY_Er9Y_eE2JfvA-xB5tGvf8Y5mwtmHaaPrFvzH0UcM9z4uaZ996u2Q6ODp3GN_f5xcrn4Vp-c_f6-OTwusFMyFI-xrSVpg3wIqq7ByrlXUWanR6U4Li02HJbm-paaivkaFdWNRO63IgdxjR1vf9eJG6jCniHYw6-hHGx9MsN68fpn8jbkO90YrWWuossG3J4MY7hZKsxl9QhoGO1FYkilVqwBqkHVGv75Bb8MSpxwvUzrPtM2DzVS5pTCGlCL1z58BYTbdmW13Jrdm_ndnNjHkVpQyPF1TfLF-R_UIWq2cpg</recordid><startdate>20211101</startdate><enddate>20211101</enddate><creator>Zhang, Da</creator><creator>Wu, Junhuang</creator><creator>Sun, Jianfei</creator><creator>Bai, Caixia</creator><creator>Xu, Fazhi</creator><creator>Duan, Pengfei</creator><creator>Wang, Yong</creator><general>Springer International Publishing</general><general>Springer Nature B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-8724-0370</orcidid></search><sort><creationdate>20211101</creationdate><title>Establishment of TaqMan-based real-time PCR assay for rapid detection of duck circovirus</title><author>Zhang, Da ; Wu, Junhuang ; Sun, Jianfei ; Bai, Caixia ; Xu, Fazhi ; Duan, Pengfei ; Wang, Yong</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c451t-becf63e80cf91c5a5c4bb95eda38cb8d80ac7dc2ebf9e74ef6c5c67ac8b85eb13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Agriculture</topic><topic>Assaying</topic><topic>Atrophy</topic><topic>Bioinformatics</topic><topic>Biomaterials</topic><topic>Biotechnology</topic><topic>Cancer Research</topic><topic>Chemistry</topic><topic>Chemistry and Materials Science</topic><topic>Cholangitis</topic><topic>Cross-reaction</topic><topic>Dwarfism</topic><topic>Epidemiology</topic><topic>Original</topic><topic>Original Article</topic><topic>Real time</topic><topic>Rep gene</topic><topic>Stem Cells</topic><topic>Waterfowl</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Da</creatorcontrib><creatorcontrib>Wu, Junhuang</creatorcontrib><creatorcontrib>Sun, Jianfei</creatorcontrib><creatorcontrib>Bai, Caixia</creatorcontrib><creatorcontrib>Xu, Fazhi</creatorcontrib><creatorcontrib>Duan, Pengfei</creatorcontrib><creatorcontrib>Wang, Yong</creatorcontrib><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>3 Biotech</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Da</au><au>Wu, Junhuang</au><au>Sun, Jianfei</au><au>Bai, Caixia</au><au>Xu, Fazhi</au><au>Duan, Pengfei</au><au>Wang, Yong</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Establishment of TaqMan-based real-time PCR assay for rapid detection of duck circovirus</atitle><jtitle>3 Biotech</jtitle><stitle>3 Biotech</stitle><date>2021-11-01</date><risdate>2021</risdate><volume>11</volume><issue>11</issue><spage>470</spage><epage>470</epage><pages>470-470</pages><artnum>470</artnum><issn>2190-572X</issn><eissn>2190-5738</eissn><abstract>Duck circovirus (DuCV) is widespread across the world and causes feather disorders in young ducks. It was identified as the causative pathogen of duck beak atrophy and dwarfism syndrome and primary sclerosing cholangitis. In this study, we aimed to establish a TaqMan-based real-time PCR assay to detect DuCV. The primers and probe were designed based on the conserved region of the DuCV Rep gene. After optimizing the reaction conditions, the minimum virus detection limit of the designed PCR technique was 39.4 copies/μL, 100 times that of conventional PCR (cPCR). No cross-reaction with six other common duck viruses was observed. The intra- and inter-assay variations were less than 1%. The detection rate of DuCV-positive clinical samples using TaqMan-based real-time PCR was higher than that using SYBR Green-based real-time PCR and cPCR. Collectively, these results showed that the established TaqMan-based real-time PCR detected DuCV with high sensitivity and specificity, and significant repeatability, making it suitable for clinical use. Hence, it may be used as a novel tool for the diagnosis and epidemiological investigation of DuCV.</abstract><cop>Cham</cop><pub>Springer International Publishing</pub><pmid>34745821</pmid><doi>10.1007/s13205-021-03021-1</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0001-8724-0370</orcidid><oa>free_for_read</oa></addata></record> |
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subjects | Agriculture Assaying Atrophy Bioinformatics Biomaterials Biotechnology Cancer Research Chemistry Chemistry and Materials Science Cholangitis Cross-reaction Dwarfism Epidemiology Original Original Article Real time Rep gene Stem Cells Waterfowl |
title | Establishment of TaqMan-based real-time PCR assay for rapid detection of duck circovirus |
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