A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China
SYBR Green coupled to melting curve analysis has been suggested to detect RNA viruses showing high genomic variability. Here, a SYBR Green-based real-time RT-PCR assay was developed for simultaneous detection and differentiation of highly pathogenic porcine reproductive and respiratory syndrome viru...
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| Veröffentlicht in: | Archives of virology 2013-02, Vol.158 (2), p.407-415 |
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| creator | Chai, Zheng Ma, Wenjun Fu, Fang Lang, Yuekun Wang, Wei Tong, Guangzhi Liu, Qinfang Cai, Xuehui Li, Xi |
| description | SYBR Green coupled to melting curve analysis has been suggested to detect RNA viruses showing high genomic variability. Here, a SYBR Green-based real-time RT-PCR assay was developed for simultaneous detection and differentiation of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) and classical type 2 PRRSV (C-PRRSV). The different strains were identified by their distinctive melting temperatures: 82.98 ± 0.25 °C and 85.95 ± 0.24 °C for HP-PRRSVs or 82.74 ± 0.26 °C for C-PRRSVs. Specificity was tested using nine other viral and bacterial pathogens of swine. The detection limit was 1 TCID
50
for HP- or C-PRRSV. Furthermore, the detection results for samples from an animal trial with HP- or C-PRRSV infections showed that the SYBR Green-based real-time RT-PCR was more sensitive than the conventional RT-PCR. Additionally, an analysis of 319 field samples from North China, Central China and Northeast China showed that HP- and C-PRRSVs co-circulated in pig herds. Thus, the SYBR Green-based real-time RT-PCR, which can be performed within one hour, is a rapid, sensitive and low-cost diagnostic tool for rapid differential detection and routine surveillance of HP- and classical type 2 PRRSVs in China. |
| doi_str_mv | 10.1007/s00705-012-1504-7 |
| format | Article |
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50
for HP- or C-PRRSV. Furthermore, the detection results for samples from an animal trial with HP- or C-PRRSV infections showed that the SYBR Green-based real-time RT-PCR was more sensitive than the conventional RT-PCR. Additionally, an analysis of 319 field samples from North China, Central China and Northeast China showed that HP- and C-PRRSVs co-circulated in pig herds. Thus, the SYBR Green-based real-time RT-PCR, which can be performed within one hour, is a rapid, sensitive and low-cost diagnostic tool for rapid differential detection and routine surveillance of HP- and classical type 2 PRRSVs in China.</description><identifier>ISSN: 0304-8608</identifier><identifier>EISSN: 1432-8798</identifier><identifier>DOI: 10.1007/s00705-012-1504-7</identifier><identifier>PMID: 23070137</identifier><language>eng</language><publisher>Vienna: Springer Vienna</publisher><subject>Animals ; Biomedical and Life Sciences ; Biomedicine ; Cell culture ; China ; Foot & mouth disease ; Genes ; Genomes ; Hogs ; Infectious Diseases ; Medical Microbiology ; Molecular Diagnostic Techniques - economics ; Molecular Diagnostic Techniques - methods ; Organic Chemicals - metabolism ; Original Article ; Pathogens ; Porcine Reproductive and Respiratory Syndrome - diagnosis ; Porcine Reproductive and Respiratory Syndrome - virology ; Porcine respiratory and reproductive syndrome virus ; Porcine respiratory and reproductive syndrome virus - genetics ; Porcine respiratory and reproductive syndrome virus - isolation & purification ; Real-Time Polymerase Chain Reaction - economics ; Real-Time Polymerase Chain Reaction - methods ; Reproducibility ; Research centers ; Sensitivity and Specificity ; Staining and Labeling - economics ; Staining and Labeling - methods ; Swine ; Time Factors ; Transition Temperature ; Veterinary medicine ; Veterinary Medicine - economics ; Veterinary Medicine - methods ; Virology ; Virology - economics ; Virology - methods ; Viruses</subject><ispartof>Archives of virology, 2013-02, Vol.158 (2), p.407-415</ispartof><rights>Springer-Verlag Wien 2012</rights><rights>Springer-Verlag Wien 2013</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c405t-d20ec637e7126972bac405fcbdf24a110e6514629e0776e4e2f8ead93745f7f53</citedby><cites>FETCH-LOGICAL-c405t-d20ec637e7126972bac405fcbdf24a110e6514629e0776e4e2f8ead93745f7f53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s00705-012-1504-7$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00705-012-1504-7$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27924,27925,41488,42557,51319</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23070137$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chai, Zheng</creatorcontrib><creatorcontrib>Ma, Wenjun</creatorcontrib><creatorcontrib>Fu, Fang</creatorcontrib><creatorcontrib>Lang, Yuekun</creatorcontrib><creatorcontrib>Wang, Wei</creatorcontrib><creatorcontrib>Tong, Guangzhi</creatorcontrib><creatorcontrib>Liu, Qinfang</creatorcontrib><creatorcontrib>Cai, Xuehui</creatorcontrib><creatorcontrib>Li, Xi</creatorcontrib><title>A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China</title><title>Archives of virology</title><addtitle>Arch Virol</addtitle><addtitle>Arch Virol</addtitle><description>SYBR Green coupled to melting curve analysis has been suggested to detect RNA viruses showing high genomic variability. Here, a SYBR Green-based real-time RT-PCR assay was developed for simultaneous detection and differentiation of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) and classical type 2 PRRSV (C-PRRSV). The different strains were identified by their distinctive melting temperatures: 82.98 ± 0.25 °C and 85.95 ± 0.24 °C for HP-PRRSVs or 82.74 ± 0.26 °C for C-PRRSVs. Specificity was tested using nine other viral and bacterial pathogens of swine. The detection limit was 1 TCID
50
for HP- or C-PRRSV. Furthermore, the detection results for samples from an animal trial with HP- or C-PRRSV infections showed that the SYBR Green-based real-time RT-PCR was more sensitive than the conventional RT-PCR. Additionally, an analysis of 319 field samples from North China, Central China and Northeast China showed that HP- and C-PRRSVs co-circulated in pig herds. Thus, the SYBR Green-based real-time RT-PCR, which can be performed within one hour, is a rapid, sensitive and low-cost diagnostic tool for rapid differential detection and routine surveillance of HP- and classical type 2 PRRSVs in China.</description><subject>Animals</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedicine</subject><subject>Cell culture</subject><subject>China</subject><subject>Foot & mouth disease</subject><subject>Genes</subject><subject>Genomes</subject><subject>Hogs</subject><subject>Infectious Diseases</subject><subject>Medical Microbiology</subject><subject>Molecular Diagnostic Techniques - economics</subject><subject>Molecular Diagnostic Techniques - methods</subject><subject>Organic Chemicals - metabolism</subject><subject>Original Article</subject><subject>Pathogens</subject><subject>Porcine Reproductive and Respiratory Syndrome - diagnosis</subject><subject>Porcine Reproductive and Respiratory Syndrome - virology</subject><subject>Porcine respiratory and reproductive syndrome virus</subject><subject>Porcine respiratory and reproductive syndrome virus - genetics</subject><subject>Porcine respiratory and reproductive syndrome virus - isolation & purification</subject><subject>Real-Time Polymerase Chain Reaction - economics</subject><subject>Real-Time Polymerase Chain Reaction - methods</subject><subject>Reproducibility</subject><subject>Research centers</subject><subject>Sensitivity and Specificity</subject><subject>Staining and Labeling - economics</subject><subject>Staining and Labeling - methods</subject><subject>Swine</subject><subject>Time Factors</subject><subject>Transition Temperature</subject><subject>Veterinary medicine</subject><subject>Veterinary Medicine - economics</subject><subject>Veterinary Medicine - methods</subject><subject>Virology</subject><subject>Virology - economics</subject><subject>Virology - methods</subject><subject>Viruses</subject><issn>0304-8608</issn><issn>1432-8798</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNks2KFDEUhQtRnHb0AdxIwI2baJL6SWo5NjoKA0o7LlwV6eSmO0NVUiZVA_WUvpK3fxQRBDcJnHz3nCSconjO2WvOmHyTcWE1ZVxQXrOKygfFileloEq26mGxYiWKqmHqoniS8x1jKJT14-JClDjIS7kqflyRL9_ebsh1Agh0qzNYkkD3dPIDkM0t_bzeEJ2zXoiLiWQ_jD0QHZDSo7fEwgRm8jEcNeudgwRh8vqoRUf2frfvFzLqaR93ELw5gqZHT290T6ZlBCLIGJPxATB7TNHOaHl_joE8-qSnmBaSl2BTxHvd-zRnYnwyc49JYUd8IOu9D_pp8cjpPsOz835ZfH3_7nb9gd58uv64vrqhpmL1RK1gYJpSguSiaaXY6oPuzNY6UWnOGTQ1rxrRApOygQqEU6BtW8qqdtLV5WXx6uSL1_0-Q566wWcDfa8DxDl3XLQcP56r8j9QJWquWM0QffkXehfnFPAhSEnVikqJFil-okyKOSdw3Zj8oNPScdYditGditFhMbpDMTqJMy_OzvN2APt74lcTEBAnIONR2EH6I_qfrj8BKrjFcg</recordid><startdate>20130201</startdate><enddate>20130201</enddate><creator>Chai, Zheng</creator><creator>Ma, Wenjun</creator><creator>Fu, Fang</creator><creator>Lang, Yuekun</creator><creator>Wang, Wei</creator><creator>Tong, Guangzhi</creator><creator>Liu, Qinfang</creator><creator>Cai, Xuehui</creator><creator>Li, Xi</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>3V.</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20130201</creationdate><title>A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China</title><author>Chai, Zheng ; Ma, Wenjun ; Fu, Fang ; Lang, Yuekun ; Wang, Wei ; Tong, Guangzhi ; Liu, Qinfang ; Cai, Xuehui ; Li, Xi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c405t-d20ec637e7126972bac405fcbdf24a110e6514629e0776e4e2f8ead93745f7f53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Animals</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedicine</topic><topic>Cell culture</topic><topic>China</topic><topic>Foot & mouth disease</topic><topic>Genes</topic><topic>Genomes</topic><topic>Hogs</topic><topic>Infectious Diseases</topic><topic>Medical Microbiology</topic><topic>Molecular Diagnostic Techniques - 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Academic</collection><jtitle>Archives of virology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chai, Zheng</au><au>Ma, Wenjun</au><au>Fu, Fang</au><au>Lang, Yuekun</au><au>Wang, Wei</au><au>Tong, Guangzhi</au><au>Liu, Qinfang</au><au>Cai, Xuehui</au><au>Li, Xi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China</atitle><jtitle>Archives of virology</jtitle><stitle>Arch Virol</stitle><addtitle>Arch Virol</addtitle><date>2013-02-01</date><risdate>2013</risdate><volume>158</volume><issue>2</issue><spage>407</spage><epage>415</epage><pages>407-415</pages><issn>0304-8608</issn><eissn>1432-8798</eissn><abstract>SYBR Green coupled to melting curve analysis has been suggested to detect RNA viruses showing high genomic variability. Here, a SYBR Green-based real-time RT-PCR assay was developed for simultaneous detection and differentiation of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) and classical type 2 PRRSV (C-PRRSV). The different strains were identified by their distinctive melting temperatures: 82.98 ± 0.25 °C and 85.95 ± 0.24 °C for HP-PRRSVs or 82.74 ± 0.26 °C for C-PRRSVs. Specificity was tested using nine other viral and bacterial pathogens of swine. The detection limit was 1 TCID
50
for HP- or C-PRRSV. Furthermore, the detection results for samples from an animal trial with HP- or C-PRRSV infections showed that the SYBR Green-based real-time RT-PCR was more sensitive than the conventional RT-PCR. Additionally, an analysis of 319 field samples from North China, Central China and Northeast China showed that HP- and C-PRRSVs co-circulated in pig herds. Thus, the SYBR Green-based real-time RT-PCR, which can be performed within one hour, is a rapid, sensitive and low-cost diagnostic tool for rapid differential detection and routine surveillance of HP- and classical type 2 PRRSVs in China.</abstract><cop>Vienna</cop><pub>Springer Vienna</pub><pmid>23070137</pmid><doi>10.1007/s00705-012-1504-7</doi><tpages>9</tpages></addata></record> |
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| ispartof | Archives of virology, 2013-02, Vol.158 (2), p.407-415 |
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| source | MEDLINE; SpringerNature Journals |
| subjects | Animals Biomedical and Life Sciences Biomedicine Cell culture China Foot & mouth disease Genes Genomes Hogs Infectious Diseases Medical Microbiology Molecular Diagnostic Techniques - economics Molecular Diagnostic Techniques - methods Organic Chemicals - metabolism Original Article Pathogens Porcine Reproductive and Respiratory Syndrome - diagnosis Porcine Reproductive and Respiratory Syndrome - virology Porcine respiratory and reproductive syndrome virus Porcine respiratory and reproductive syndrome virus - genetics Porcine respiratory and reproductive syndrome virus - isolation & purification Real-Time Polymerase Chain Reaction - economics Real-Time Polymerase Chain Reaction - methods Reproducibility Research centers Sensitivity and Specificity Staining and Labeling - economics Staining and Labeling - methods Swine Time Factors Transition Temperature Veterinary medicine Veterinary Medicine - economics Veterinary Medicine - methods Virology Virology - economics Virology - methods Viruses |
| title | A SYBR Green-based real-time RT-PCR assay for simple and rapid detection and differentiation of highly pathogenic and classical type 2 porcine reproductive and respiratory syndrome virus circulating in China |
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