Real-Time PCR Detection of Parapoxvirus DNA
Detection of parapoxviruses is important in various animals as well as in humans as zoonotic infections. Reliable detection of parapoxviruses is fundamental for the exclusion of other rash-causing illnesses, for both veterinarians and medical practitioners. To date, however, no real-time PCR assay f...
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| Veröffentlicht in: | Clinical chemistry (Baltimore, Md.) Md.), 2006-02, Vol.52 (2), p.316-319 |
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| creator | Nitsche, Andreas Buttner, Mathias Wilhelm, Sonja Pauli, Georg Meyer, Hermann |
| description | Detection of parapoxviruses is important in various animals as well as in humans as zoonotic infections. Reliable detection of parapoxviruses is fundamental for the exclusion of other rash-causing illnesses, for both veterinarians and medical practitioners. To date, however, no real-time PCR assay for the detection of parapoxviruses has been reported.
A minor groove binder-based quantitative real-time PCR assay targeting the B2L gene of parapoxviruses was developed on the ABI Prism and the LightCycler platforms.
The real-time PCR assay successfully amplified DNA fragments from a total of 41 parapoxvirus strains and isolates representing the species orf virus, bovine papular stomatitis virus, pseudocowpoxvirus, and sealpoxvirus. Probit analysis gave a limit of detection of 4.7 copies per assay (95% confidence interval, 3.7-6.8 copies per reaction). Scabs contain a sufficient amount of parapoxvirus DNA and can therefore be used for PCR without any DNA preparation step. No cross-reactivity to human, bovine, or sheep genomic DNA or other DNA viruses, including orthopoxviruses, molluscum contagiosum viruses, and yaba-like disease viruses, was observed.
The presented assay is suitable for the detection of parapoxvirus infections in clinical material of human and animal origin. |
| doi_str_mv | 10.1373/clinchem.2005.060335 |
| format | Article |
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A minor groove binder-based quantitative real-time PCR assay targeting the B2L gene of parapoxviruses was developed on the ABI Prism and the LightCycler platforms.
The real-time PCR assay successfully amplified DNA fragments from a total of 41 parapoxvirus strains and isolates representing the species orf virus, bovine papular stomatitis virus, pseudocowpoxvirus, and sealpoxvirus. Probit analysis gave a limit of detection of 4.7 copies per assay (95% confidence interval, 3.7-6.8 copies per reaction). Scabs contain a sufficient amount of parapoxvirus DNA and can therefore be used for PCR without any DNA preparation step. No cross-reactivity to human, bovine, or sheep genomic DNA or other DNA viruses, including orthopoxviruses, molluscum contagiosum viruses, and yaba-like disease viruses, was observed.
The presented assay is suitable for the detection of parapoxvirus infections in clinical material of human and animal origin.</description><identifier>ISSN: 0009-9147</identifier><identifier>EISSN: 1530-8561</identifier><identifier>DOI: 10.1373/clinchem.2005.060335</identifier><identifier>PMID: 16449215</identifier><identifier>CODEN: CLCHAU</identifier><language>eng</language><publisher>Washington, DC: Am Assoc Clin Chem</publisher><subject>Analytical, structural and metabolic biochemistry ; Animals ; Biological and medical sciences ; Bovine papular stomatitis virus ; Confidence intervals ; Deoxyribonucleic acid ; DNA ; DNA, Viral - analysis ; Fundamental and applied biological sciences. Psychology ; Humans ; Infections ; Investigative techniques, diagnostic techniques (general aspects) ; Medical sciences ; Mollusca ; Mollusks ; Orf virus ; Parapoxvirus ; Parapoxvirus - genetics ; Parapoxvirus - isolation & purification ; Plasmids ; Poxviridae Infections - veterinary ; Poxviridae Infections - virology ; Reverse Transcriptase Polymerase Chain Reaction - methods ; Reverse Transcriptase Polymerase Chain Reaction - veterinary ; Sensitivity and Specificity ; Sheep ; Viruses</subject><ispartof>Clinical chemistry (Baltimore, Md.), 2006-02, Vol.52 (2), p.316-319</ispartof><rights>2006 INIST-CNRS</rights><rights>Copyright American Association for Clinical Chemistry Feb 2006</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c535t-633616a1e9640cb51c42cf596c6f2b4a3dbc22edffa51f254ee5a2867f6d49943</citedby><cites>FETCH-LOGICAL-c535t-633616a1e9640cb51c42cf596c6f2b4a3dbc22edffa51f254ee5a2867f6d49943</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17594687$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16449215$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Nitsche, Andreas</creatorcontrib><creatorcontrib>Buttner, Mathias</creatorcontrib><creatorcontrib>Wilhelm, Sonja</creatorcontrib><creatorcontrib>Pauli, Georg</creatorcontrib><creatorcontrib>Meyer, Hermann</creatorcontrib><title>Real-Time PCR Detection of Parapoxvirus DNA</title><title>Clinical chemistry (Baltimore, Md.)</title><addtitle>Clin Chem</addtitle><description>Detection of parapoxviruses is important in various animals as well as in humans as zoonotic infections. Reliable detection of parapoxviruses is fundamental for the exclusion of other rash-causing illnesses, for both veterinarians and medical practitioners. To date, however, no real-time PCR assay for the detection of parapoxviruses has been reported.
A minor groove binder-based quantitative real-time PCR assay targeting the B2L gene of parapoxviruses was developed on the ABI Prism and the LightCycler platforms.
The real-time PCR assay successfully amplified DNA fragments from a total of 41 parapoxvirus strains and isolates representing the species orf virus, bovine papular stomatitis virus, pseudocowpoxvirus, and sealpoxvirus. Probit analysis gave a limit of detection of 4.7 copies per assay (95% confidence interval, 3.7-6.8 copies per reaction). Scabs contain a sufficient amount of parapoxvirus DNA and can therefore be used for PCR without any DNA preparation step. No cross-reactivity to human, bovine, or sheep genomic DNA or other DNA viruses, including orthopoxviruses, molluscum contagiosum viruses, and yaba-like disease viruses, was observed.
The presented assay is suitable for the detection of parapoxvirus infections in clinical material of human and animal origin.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Bovine papular stomatitis virus</subject><subject>Confidence intervals</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA, Viral - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Infections</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Medical sciences</subject><subject>Mollusca</subject><subject>Mollusks</subject><subject>Orf virus</subject><subject>Parapoxvirus</subject><subject>Parapoxvirus - genetics</subject><subject>Parapoxvirus - isolation & purification</subject><subject>Plasmids</subject><subject>Poxviridae Infections - veterinary</subject><subject>Poxviridae Infections - virology</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - methods</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - veterinary</subject><subject>Sensitivity and Specificity</subject><subject>Sheep</subject><subject>Viruses</subject><issn>0009-9147</issn><issn>1530-8561</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqFkNtKAzEQhoMoWg9vILIIeiNbMzluLqUeQVSKXoc0TWxkDzXpWn17t3RF8MarYeCbf2Y-hA4BD4FKem7LUNuZq4YEYz7EAlPKN9AAOMV5wQVsogHGWOUKmNxBuym9dS2ThdhGOyAYUwT4AJ2NnSnz51C57Gk0zi7dwtlFaOqs8dmTiWbefH6E2Kbs8uFiH215UyZ30Nc99HJ99Ty6ze8fb-5GF_e55ZQvckGpAGHAKcGwnXCwjFjPlbDCkwkzdDqxhLip94aDJ5w5xw0phPRiypRidA-drnPnsXlvXVroKiTrytLUrmmTllhS3u34FwRV0EIUsgOP_4BvTRvr7glNgGHAQvIOYmvIxial6Lyex1CZ-KUB65Vy_aNcr5TrtfJu7KjPbieVm_4O9Y474KQHTLKm9NHUNqRfTnLF1kf23Cy8zpYhOp0qU5ZdLOjlcsmJJpqCoN-j5pVo</recordid><startdate>20060201</startdate><enddate>20060201</enddate><creator>Nitsche, Andreas</creator><creator>Buttner, Mathias</creator><creator>Wilhelm, Sonja</creator><creator>Pauli, Georg</creator><creator>Meyer, Hermann</creator><general>Am Assoc Clin Chem</general><general>American Association for Clinical Chemistry</general><general>Oxford University Press</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>3V.</scope><scope>4U-</scope><scope>7QO</scope><scope>7RV</scope><scope>7TM</scope><scope>7U7</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PCBAR</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>S0X</scope><scope>7T7</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20060201</creationdate><title>Real-Time PCR Detection of Parapoxvirus DNA</title><author>Nitsche, Andreas ; Buttner, Mathias ; Wilhelm, Sonja ; Pauli, Georg ; Meyer, Hermann</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c535t-633616a1e9640cb51c42cf596c6f2b4a3dbc22edffa51f254ee5a2867f6d49943</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Bovine papular stomatitis virus</topic><topic>Confidence intervals</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA, Viral - analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Infections</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Medical sciences</topic><topic>Mollusca</topic><topic>Mollusks</topic><topic>Orf virus</topic><topic>Parapoxvirus</topic><topic>Parapoxvirus - genetics</topic><topic>Parapoxvirus - isolation & purification</topic><topic>Plasmids</topic><topic>Poxviridae Infections - veterinary</topic><topic>Poxviridae Infections - virology</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - methods</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - veterinary</topic><topic>Sensitivity and Specificity</topic><topic>Sheep</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Nitsche, Andreas</creatorcontrib><creatorcontrib>Buttner, Mathias</creatorcontrib><creatorcontrib>Wilhelm, Sonja</creatorcontrib><creatorcontrib>Pauli, Georg</creatorcontrib><creatorcontrib>Meyer, Hermann</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>ProQuest Central (Corporate)</collection><collection>University Readers</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>Materials Science Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>SIRS Editorial</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Nitsche, Andreas</au><au>Buttner, Mathias</au><au>Wilhelm, Sonja</au><au>Pauli, Georg</au><au>Meyer, Hermann</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Real-Time PCR Detection of Parapoxvirus DNA</atitle><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle><addtitle>Clin Chem</addtitle><date>2006-02-01</date><risdate>2006</risdate><volume>52</volume><issue>2</issue><spage>316</spage><epage>319</epage><pages>316-319</pages><issn>0009-9147</issn><eissn>1530-8561</eissn><coden>CLCHAU</coden><abstract>Detection of parapoxviruses is important in various animals as well as in humans as zoonotic infections. Reliable detection of parapoxviruses is fundamental for the exclusion of other rash-causing illnesses, for both veterinarians and medical practitioners. To date, however, no real-time PCR assay for the detection of parapoxviruses has been reported.
A minor groove binder-based quantitative real-time PCR assay targeting the B2L gene of parapoxviruses was developed on the ABI Prism and the LightCycler platforms.
The real-time PCR assay successfully amplified DNA fragments from a total of 41 parapoxvirus strains and isolates representing the species orf virus, bovine papular stomatitis virus, pseudocowpoxvirus, and sealpoxvirus. Probit analysis gave a limit of detection of 4.7 copies per assay (95% confidence interval, 3.7-6.8 copies per reaction). Scabs contain a sufficient amount of parapoxvirus DNA and can therefore be used for PCR without any DNA preparation step. No cross-reactivity to human, bovine, or sheep genomic DNA or other DNA viruses, including orthopoxviruses, molluscum contagiosum viruses, and yaba-like disease viruses, was observed.
The presented assay is suitable for the detection of parapoxvirus infections in clinical material of human and animal origin.</abstract><cop>Washington, DC</cop><pub>Am Assoc Clin Chem</pub><pmid>16449215</pmid><doi>10.1373/clinchem.2005.060335</doi><tpages>4</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Clinical chemistry (Baltimore, Md.), 2006-02, Vol.52 (2), p.316-319 |
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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE |
| subjects | Analytical, structural and metabolic biochemistry Animals Biological and medical sciences Bovine papular stomatitis virus Confidence intervals Deoxyribonucleic acid DNA DNA, Viral - analysis Fundamental and applied biological sciences. Psychology Humans Infections Investigative techniques, diagnostic techniques (general aspects) Medical sciences Mollusca Mollusks Orf virus Parapoxvirus Parapoxvirus - genetics Parapoxvirus - isolation & purification Plasmids Poxviridae Infections - veterinary Poxviridae Infections - virology Reverse Transcriptase Polymerase Chain Reaction - methods Reverse Transcriptase Polymerase Chain Reaction - veterinary Sensitivity and Specificity Sheep Viruses |
| title | Real-Time PCR Detection of Parapoxvirus DNA |
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