Rapid detection of goose hemorrhagic polyomavirus using TaqMan quantitative real-time PCR
Due to low doses of infection, an efficient and sensitive virus detection method is necessary to detect low amounts of goose hemorrhagic polyomavirus (GHPV). In this study, we have developed a TaqMan real-time PCR (qPCR) specific assay for the detection of GHPV. Specificity assay showed no cross-rea...
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Veröffentlicht in: | Molecular and cellular probes 2018-06, Vol.39, p.61-64 |
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creator | Wan, Chunhe Cheng, Longfei Fu, Guanghua Chen, Cuiteng Liu, Rongchang Shi, Shaohua Chen, Hongmei Fu, Qiuling Huang, Yu |
description | Due to low doses of infection, an efficient and sensitive virus detection method is necessary to detect low amounts of goose hemorrhagic polyomavirus (GHPV). In this study, we have developed a TaqMan real-time PCR (qPCR) specific assay for the detection of GHPV. Specificity assay showed no cross-reactions with other common waterfowl viruses. The standard curve had a linear correlation of 0.997 and efficiency of 99% between the cycle threshold value and the logarithm of the plasmids copy number. The possible lowest detectable concentration was 35.4 copies/μl; 100 times more sensitive than conventional PCR (detection limit, 3.54 × 103 copies/μl). Domestic Jinyun Sheldrakes ducks and their embryonated eggs were found positive of GHPV infection which provides evidence of possible vertical transmission of GHPV.
•A TaqMan real-time PCR method for goose hemorrhagic polyomavirus (GHPV) was developed.•The method showed high sensitivity, specificity and reproducibility.•Domestic Sheldrakes can be infected in GHPV, which indicates GHPV persistent infection in ducks, China. |
doi_str_mv | 10.1016/j.mcp.2018.04.003 |
format | Article |
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•A TaqMan real-time PCR method for goose hemorrhagic polyomavirus (GHPV) was developed.•The method showed high sensitivity, specificity and reproducibility.•Domestic Sheldrakes can be infected in GHPV, which indicates GHPV persistent infection in ducks, China.</description><identifier>ISSN: 0890-8508</identifier><identifier>EISSN: 1096-1194</identifier><identifier>DOI: 10.1016/j.mcp.2018.04.003</identifier><identifier>PMID: 29665412</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>Animals ; Domestic Jinyun Sheldrakes ; Geese - virology ; Goose hemorrhagic polyomavirus ; Polyomavirus - isolation & purification ; Real-Time Polymerase Chain Reaction - methods ; Reproducibility of Results ; Sensitivity ; Sensitivity and Specificity ; TaqMan real-time PCR</subject><ispartof>Molecular and cellular probes, 2018-06, Vol.39, p.61-64</ispartof><rights>2018</rights><rights>Copyright © 2018. Published by Elsevier Ltd.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c419t-d993f20904d8e32e9be6558111727cc4f790f5b1c8abed12bca005521c0d33b83</citedby><cites>FETCH-LOGICAL-c419t-d993f20904d8e32e9be6558111727cc4f790f5b1c8abed12bca005521c0d33b83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.mcp.2018.04.003$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29665412$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wan, Chunhe</creatorcontrib><creatorcontrib>Cheng, Longfei</creatorcontrib><creatorcontrib>Fu, Guanghua</creatorcontrib><creatorcontrib>Chen, Cuiteng</creatorcontrib><creatorcontrib>Liu, Rongchang</creatorcontrib><creatorcontrib>Shi, Shaohua</creatorcontrib><creatorcontrib>Chen, Hongmei</creatorcontrib><creatorcontrib>Fu, Qiuling</creatorcontrib><creatorcontrib>Huang, Yu</creatorcontrib><title>Rapid detection of goose hemorrhagic polyomavirus using TaqMan quantitative real-time PCR</title><title>Molecular and cellular probes</title><addtitle>Mol Cell Probes</addtitle><description>Due to low doses of infection, an efficient and sensitive virus detection method is necessary to detect low amounts of goose hemorrhagic polyomavirus (GHPV). In this study, we have developed a TaqMan real-time PCR (qPCR) specific assay for the detection of GHPV. Specificity assay showed no cross-reactions with other common waterfowl viruses. The standard curve had a linear correlation of 0.997 and efficiency of 99% between the cycle threshold value and the logarithm of the plasmids copy number. The possible lowest detectable concentration was 35.4 copies/μl; 100 times more sensitive than conventional PCR (detection limit, 3.54 × 103 copies/μl). Domestic Jinyun Sheldrakes ducks and their embryonated eggs were found positive of GHPV infection which provides evidence of possible vertical transmission of GHPV.
•A TaqMan real-time PCR method for goose hemorrhagic polyomavirus (GHPV) was developed.•The method showed high sensitivity, specificity and reproducibility.•Domestic Sheldrakes can be infected in GHPV, which indicates GHPV persistent infection in ducks, China.</description><subject>Animals</subject><subject>Domestic Jinyun Sheldrakes</subject><subject>Geese - virology</subject><subject>Goose hemorrhagic polyomavirus</subject><subject>Polyomavirus - isolation & purification</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Reproducibility of Results</subject><subject>Sensitivity</subject><subject>Sensitivity and Specificity</subject><subject>TaqMan real-time PCR</subject><issn>0890-8508</issn><issn>1096-1194</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE9v1DAQRy1ERZfCB-CCfOSSMOP8s8UJrQpUKmpVlQMny7EnW6-SOGsnK_XbN9UWjj3N5f2eNI-xTwg5AtZf9_lgp1wAyhzKHKB4wzYIqs4QVfmWbUAqyGQF8py9T2kPAKoE-Y6dC1XXVYliw_7emck77mgmO_sw8tDxXQiJ-AMNIcYHs_OWT6F_DIM5-rgkviQ_7vi9Ofw2Iz8sZpz9bGZ_JB7J9NnsB-K327sP7KwzfaKPL_eC_flxeb_9lV3f_Lzafr_ObIlqzpxSRSdAQekkFYJUS3VVSURsRGNt2TUKuqpFK01LDkVrDUBVCbTgiqKVxQX7cvJOMRwWSrMefLLU92aksCQtQDRQN7UQK4on1MaQUqROT9EPJj5qBP1cVO_1WlQ_F9VQ6rXouvn8ol_agdz_xb-EK_DtBND65NFT1Ml6Gi05H9em2gX_iv4JY0qGzA</recordid><startdate>201806</startdate><enddate>201806</enddate><creator>Wan, Chunhe</creator><creator>Cheng, Longfei</creator><creator>Fu, Guanghua</creator><creator>Chen, Cuiteng</creator><creator>Liu, Rongchang</creator><creator>Shi, Shaohua</creator><creator>Chen, Hongmei</creator><creator>Fu, Qiuling</creator><creator>Huang, Yu</creator><general>Elsevier Ltd</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>201806</creationdate><title>Rapid detection of goose hemorrhagic polyomavirus using TaqMan quantitative real-time PCR</title><author>Wan, Chunhe ; Cheng, Longfei ; Fu, Guanghua ; Chen, Cuiteng ; Liu, Rongchang ; Shi, Shaohua ; Chen, Hongmei ; Fu, Qiuling ; Huang, Yu</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c419t-d993f20904d8e32e9be6558111727cc4f790f5b1c8abed12bca005521c0d33b83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Animals</topic><topic>Domestic Jinyun Sheldrakes</topic><topic>Geese - virology</topic><topic>Goose hemorrhagic polyomavirus</topic><topic>Polyomavirus - isolation & purification</topic><topic>Real-Time Polymerase Chain Reaction - methods</topic><topic>Reproducibility of Results</topic><topic>Sensitivity</topic><topic>Sensitivity and Specificity</topic><topic>TaqMan real-time PCR</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wan, Chunhe</creatorcontrib><creatorcontrib>Cheng, Longfei</creatorcontrib><creatorcontrib>Fu, Guanghua</creatorcontrib><creatorcontrib>Chen, Cuiteng</creatorcontrib><creatorcontrib>Liu, Rongchang</creatorcontrib><creatorcontrib>Shi, Shaohua</creatorcontrib><creatorcontrib>Chen, Hongmei</creatorcontrib><creatorcontrib>Fu, Qiuling</creatorcontrib><creatorcontrib>Huang, Yu</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>Molecular and cellular probes</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wan, Chunhe</au><au>Cheng, Longfei</au><au>Fu, Guanghua</au><au>Chen, Cuiteng</au><au>Liu, Rongchang</au><au>Shi, Shaohua</au><au>Chen, Hongmei</au><au>Fu, Qiuling</au><au>Huang, Yu</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Rapid detection of goose hemorrhagic polyomavirus using TaqMan quantitative real-time PCR</atitle><jtitle>Molecular and cellular probes</jtitle><addtitle>Mol Cell Probes</addtitle><date>2018-06</date><risdate>2018</risdate><volume>39</volume><spage>61</spage><epage>64</epage><pages>61-64</pages><issn>0890-8508</issn><eissn>1096-1194</eissn><abstract>Due to low doses of infection, an efficient and sensitive virus detection method is necessary to detect low amounts of goose hemorrhagic polyomavirus (GHPV). In this study, we have developed a TaqMan real-time PCR (qPCR) specific assay for the detection of GHPV. Specificity assay showed no cross-reactions with other common waterfowl viruses. The standard curve had a linear correlation of 0.997 and efficiency of 99% between the cycle threshold value and the logarithm of the plasmids copy number. The possible lowest detectable concentration was 35.4 copies/μl; 100 times more sensitive than conventional PCR (detection limit, 3.54 × 103 copies/μl). Domestic Jinyun Sheldrakes ducks and their embryonated eggs were found positive of GHPV infection which provides evidence of possible vertical transmission of GHPV.
•A TaqMan real-time PCR method for goose hemorrhagic polyomavirus (GHPV) was developed.•The method showed high sensitivity, specificity and reproducibility.•Domestic Sheldrakes can be infected in GHPV, which indicates GHPV persistent infection in ducks, China.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>29665412</pmid><doi>10.1016/j.mcp.2018.04.003</doi><tpages>4</tpages></addata></record> |
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subjects | Animals Domestic Jinyun Sheldrakes Geese - virology Goose hemorrhagic polyomavirus Polyomavirus - isolation & purification Real-Time Polymerase Chain Reaction - methods Reproducibility of Results Sensitivity Sensitivity and Specificity TaqMan real-time PCR |
title | Rapid detection of goose hemorrhagic polyomavirus using TaqMan quantitative real-time PCR |
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