Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay
Spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are three important fish rhabdoviruses, causing serious Office International des Epizooties (OIE) classified diseases in wild and farmed fish. Here, a new multiplex real-...
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| Veröffentlicht in: | Journal of virological methods 2008-04, Vol.149 (1), p.103-109 |
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| creator | Liu, Zongxiao Teng, Yong Liu, Hong Jiang, Yulin Xie, Xiayang Li, Huifang Lv, Jiangqiang Gao, Longying He, Junqiang Shi, Xiujie Tian, Feiyan Yang, Jingshun Xie, Congxin |
| description | Spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are three important fish rhabdoviruses, causing serious Office International des Epizooties (OIE) classified diseases in wild and farmed fish. Here, a new multiplex real-time quantitative RT-PCR (mqRT-PCR) assay was developed for simultaneous detection, identification and quantification of these three rhabdoviruses. The sets of primers and probes were targeted to conserved regions of glycoprotein (G) gene of SVCV, nucleoprotein (N) gene of IHNV and G gene of VHSV and used to amplify. The sensitivity, specificity and interference test of mqRT-PCR assay was analyzed. It was shown that the detection levels of 100 copies of SVCV, 220 copies of IHNV and 140 copies of VHSV were achieved, and there was no non-specific amplification and cross-reactivity using RNA of pike fry rhabdovirus (PFRV), infectious pancreatic necrosis virus (IPNV) and grass carp reovirus (GCRV). A total of 80 clinical fish samples were tested using the mqRT-PCR assay and the results were confirmed by antigen-capture ELISA and cell culture assay. This assay has the potential to be used for both research applications and diagnosis. |
| doi_str_mv | 10.1016/j.jviromet.2007.12.017 |
| format | Article |
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Here, a new multiplex real-time quantitative RT-PCR (mqRT-PCR) assay was developed for simultaneous detection, identification and quantification of these three rhabdoviruses. The sets of primers and probes were targeted to conserved regions of glycoprotein (G) gene of SVCV, nucleoprotein (N) gene of IHNV and G gene of VHSV and used to amplify. The sensitivity, specificity and interference test of mqRT-PCR assay was analyzed. It was shown that the detection levels of 100 copies of SVCV, 220 copies of IHNV and 140 copies of VHSV were achieved, and there was no non-specific amplification and cross-reactivity using RNA of pike fry rhabdovirus (PFRV), infectious pancreatic necrosis virus (IPNV) and grass carp reovirus (GCRV). A total of 80 clinical fish samples were tested using the mqRT-PCR assay and the results were confirmed by antigen-capture ELISA and cell culture assay. This assay has the potential to be used for both research applications and diagnosis.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/j.jviromet.2007.12.017</identifier><identifier>PMID: 18299154</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Animals ; Biological and medical sciences ; Cell Line ; Fish Diseases - diagnosis ; Fish Diseases - virology ; Fish rhabdoviruses ; Fishes - virology ; Fundamental and applied biological sciences. Psychology ; Grass carp reovirus ; Infectious hematopoietic necrosis virus ; Infectious hematopoietic necrosis virus - isolation & purification ; Infectious pancreatic necrosis virus ; Microbiology ; mqRT-PCR ; Novirhabdovirus - isolation & purification ; Pike fry rhabdovirus ; Reverse Transcriptase Polymerase Chain Reaction - methods ; Rhabdoviridae Infections - diagnosis ; Rhabdoviridae Infections - veterinary ; Rhabdoviridae Infections - virology ; Rhabdovirus ; Sensitivity and Specificity ; Simultaneous detection ; Spring viremia of carp virus ; Techniques used in virology ; Vesiculovirus - isolation & purification ; Viral hemorrhagic septicemia virus ; Virology</subject><ispartof>Journal of virological methods, 2008-04, Vol.149 (1), p.103-109</ispartof><rights>2008 Elsevier B.V.</rights><rights>2008 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c427t-31ec550c6566da1c693394526a0315e66fa2639392905af7899d3869e75a59f83</citedby><cites>FETCH-LOGICAL-c427t-31ec550c6566da1c693394526a0315e66fa2639392905af7899d3869e75a59f83</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.jviromet.2007.12.017$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20233528$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18299154$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Liu, Zongxiao</creatorcontrib><creatorcontrib>Teng, Yong</creatorcontrib><creatorcontrib>Liu, Hong</creatorcontrib><creatorcontrib>Jiang, Yulin</creatorcontrib><creatorcontrib>Xie, Xiayang</creatorcontrib><creatorcontrib>Li, Huifang</creatorcontrib><creatorcontrib>Lv, Jiangqiang</creatorcontrib><creatorcontrib>Gao, Longying</creatorcontrib><creatorcontrib>He, Junqiang</creatorcontrib><creatorcontrib>Shi, Xiujie</creatorcontrib><creatorcontrib>Tian, Feiyan</creatorcontrib><creatorcontrib>Yang, Jingshun</creatorcontrib><creatorcontrib>Xie, Congxin</creatorcontrib><title>Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>Spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are three important fish rhabdoviruses, causing serious Office International des Epizooties (OIE) classified diseases in wild and farmed fish. Here, a new multiplex real-time quantitative RT-PCR (mqRT-PCR) assay was developed for simultaneous detection, identification and quantification of these three rhabdoviruses. The sets of primers and probes were targeted to conserved regions of glycoprotein (G) gene of SVCV, nucleoprotein (N) gene of IHNV and G gene of VHSV and used to amplify. The sensitivity, specificity and interference test of mqRT-PCR assay was analyzed. It was shown that the detection levels of 100 copies of SVCV, 220 copies of IHNV and 140 copies of VHSV were achieved, and there was no non-specific amplification and cross-reactivity using RNA of pike fry rhabdovirus (PFRV), infectious pancreatic necrosis virus (IPNV) and grass carp reovirus (GCRV). A total of 80 clinical fish samples were tested using the mqRT-PCR assay and the results were confirmed by antigen-capture ELISA and cell culture assay. This assay has the potential to be used for both research applications and diagnosis.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Fish Diseases - diagnosis</subject><subject>Fish Diseases - virology</subject><subject>Fish rhabdoviruses</subject><subject>Fishes - virology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Grass carp reovirus</subject><subject>Infectious hematopoietic necrosis virus</subject><subject>Infectious hematopoietic necrosis virus - isolation & purification</subject><subject>Infectious pancreatic necrosis virus</subject><subject>Microbiology</subject><subject>mqRT-PCR</subject><subject>Novirhabdovirus - isolation & purification</subject><subject>Pike fry rhabdovirus</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - methods</subject><subject>Rhabdoviridae Infections - diagnosis</subject><subject>Rhabdoviridae Infections - veterinary</subject><subject>Rhabdoviridae Infections - virology</subject><subject>Rhabdovirus</subject><subject>Sensitivity and Specificity</subject><subject>Simultaneous detection</subject><subject>Spring viremia of carp virus</subject><subject>Techniques used in virology</subject><subject>Vesiculovirus - isolation & purification</subject><subject>Viral hemorrhagic septicemia virus</subject><subject>Virology</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkUtvGyEURlHVqnGT_IWITbubKY8Bhl0qK31IkVql6RoR5hJjzcMBxqr_fRnZSZdZweJ8XO53ELqipKaEys_bersPcRog14wQVVNWE6reoBVtla6Ibpu3aFVAWe68OUMfUtoSQoTi_D06oy3TmopmhfzvMMx9tiNMc8IdZHA5TCOePM6bCIB9SBscN_ahm8rAOUHCcwrjI15iYdfDXxzB9lUOA-Cn2Y45ZJvDHvDdffVrfYdtSvZwgd552ye4PJ3n6M_Xm_v19-r257cf6y-3lWuYyhWn4IQgTgopO0ud1JzrRjBpCacCpPSWSa65ZpoI61WrdcdbqUEJK7Rv-Tn6dHx3F6enGVI2Q0gO-v64oFGkoZyWSl4DGVGKS7GA8gi6OKUUwZtdDIONB0OJWVSYrXlWYRYVhjJTVJTg1WnC_DBA9z926r4AH0-ATc72PtrRhfTCMcI4F2zZ6frIQSluHyCa5AKMDroQiy3TTeG1v_wDaYOrPQ</recordid><startdate>20080401</startdate><enddate>20080401</enddate><creator>Liu, Zongxiao</creator><creator>Teng, Yong</creator><creator>Liu, Hong</creator><creator>Jiang, Yulin</creator><creator>Xie, Xiayang</creator><creator>Li, Huifang</creator><creator>Lv, Jiangqiang</creator><creator>Gao, Longying</creator><creator>He, Junqiang</creator><creator>Shi, Xiujie</creator><creator>Tian, Feiyan</creator><creator>Yang, Jingshun</creator><creator>Xie, Congxin</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><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>7QL</scope><scope>7U9</scope><scope>C1K</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20080401</creationdate><title>Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay</title><author>Liu, Zongxiao ; Teng, Yong ; Liu, Hong ; Jiang, Yulin ; Xie, Xiayang ; Li, Huifang ; Lv, Jiangqiang ; Gao, Longying ; He, Junqiang ; Shi, Xiujie ; Tian, Feiyan ; Yang, Jingshun ; Xie, Congxin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c427t-31ec550c6566da1c693394526a0315e66fa2639392905af7899d3869e75a59f83</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cell Line</topic><topic>Fish Diseases - diagnosis</topic><topic>Fish Diseases - virology</topic><topic>Fish rhabdoviruses</topic><topic>Fishes - virology</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Grass carp reovirus</topic><topic>Infectious hematopoietic necrosis virus</topic><topic>Infectious hematopoietic necrosis virus - isolation & purification</topic><topic>Infectious pancreatic necrosis virus</topic><topic>Microbiology</topic><topic>mqRT-PCR</topic><topic>Novirhabdovirus - isolation & purification</topic><topic>Pike fry rhabdovirus</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - methods</topic><topic>Rhabdoviridae Infections - diagnosis</topic><topic>Rhabdoviridae Infections - veterinary</topic><topic>Rhabdoviridae Infections - virology</topic><topic>Rhabdovirus</topic><topic>Sensitivity and Specificity</topic><topic>Simultaneous detection</topic><topic>Spring viremia of carp virus</topic><topic>Techniques used in virology</topic><topic>Vesiculovirus - isolation & purification</topic><topic>Viral hemorrhagic septicemia virus</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Liu, Zongxiao</creatorcontrib><creatorcontrib>Teng, Yong</creatorcontrib><creatorcontrib>Liu, Hong</creatorcontrib><creatorcontrib>Jiang, Yulin</creatorcontrib><creatorcontrib>Xie, Xiayang</creatorcontrib><creatorcontrib>Li, Huifang</creatorcontrib><creatorcontrib>Lv, Jiangqiang</creatorcontrib><creatorcontrib>Gao, Longying</creatorcontrib><creatorcontrib>He, Junqiang</creatorcontrib><creatorcontrib>Shi, Xiujie</creatorcontrib><creatorcontrib>Tian, Feiyan</creatorcontrib><creatorcontrib>Yang, Jingshun</creatorcontrib><creatorcontrib>Xie, Congxin</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Virology and AIDS Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Liu, Zongxiao</au><au>Teng, Yong</au><au>Liu, Hong</au><au>Jiang, Yulin</au><au>Xie, Xiayang</au><au>Li, Huifang</au><au>Lv, Jiangqiang</au><au>Gao, Longying</au><au>He, Junqiang</au><au>Shi, Xiujie</au><au>Tian, Feiyan</au><au>Yang, Jingshun</au><au>Xie, Congxin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>2008-04-01</date><risdate>2008</risdate><volume>149</volume><issue>1</issue><spage>103</spage><epage>109</epage><pages>103-109</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>Spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are three important fish rhabdoviruses, causing serious Office International des Epizooties (OIE) classified diseases in wild and farmed fish. Here, a new multiplex real-time quantitative RT-PCR (mqRT-PCR) assay was developed for simultaneous detection, identification and quantification of these three rhabdoviruses. The sets of primers and probes were targeted to conserved regions of glycoprotein (G) gene of SVCV, nucleoprotein (N) gene of IHNV and G gene of VHSV and used to amplify. The sensitivity, specificity and interference test of mqRT-PCR assay was analyzed. It was shown that the detection levels of 100 copies of SVCV, 220 copies of IHNV and 140 copies of VHSV were achieved, and there was no non-specific amplification and cross-reactivity using RNA of pike fry rhabdovirus (PFRV), infectious pancreatic necrosis virus (IPNV) and grass carp reovirus (GCRV). A total of 80 clinical fish samples were tested using the mqRT-PCR assay and the results were confirmed by antigen-capture ELISA and cell culture assay. This assay has the potential to be used for both research applications and diagnosis.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>18299154</pmid><doi>10.1016/j.jviromet.2007.12.017</doi><tpages>7</tpages></addata></record> |
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| subjects | Animals Biological and medical sciences Cell Line Fish Diseases - diagnosis Fish Diseases - virology Fish rhabdoviruses Fishes - virology Fundamental and applied biological sciences. Psychology Grass carp reovirus Infectious hematopoietic necrosis virus Infectious hematopoietic necrosis virus - isolation & purification Infectious pancreatic necrosis virus Microbiology mqRT-PCR Novirhabdovirus - isolation & purification Pike fry rhabdovirus Reverse Transcriptase Polymerase Chain Reaction - methods Rhabdoviridae Infections - diagnosis Rhabdoviridae Infections - veterinary Rhabdoviridae Infections - virology Rhabdovirus Sensitivity and Specificity Simultaneous detection Spring viremia of carp virus Techniques used in virology Vesiculovirus - isolation & purification Viral hemorrhagic septicemia virus Virology |
| title | Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay |
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