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Development of a novel TaqMan-based real-time PCR assay for the detection of porcine boca-like virus (Pbo-likeV)
The recently discovered porcine boca-like virus (Pbo-likeV) is a member of the Parvoviridae family, genus Bocavirus, and it is potentially associated with swine disease. Several studies have associated Pbo-likeV with postweaning multisystemic wasting syndrome in pigs, but the full spectrum of clinic...
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Published in: | Virology journal 2011-07, Vol.8 (1), p.357-357, Article 357 |
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creator | Li, Bin Xiao, Shaobo Ma, Junjie Liu, Yanling Mao, Li Wen, Libin Mao, Aihua Zhang, Xuehan Ni, Yanxiu Guo, Rongli Zhou, Junming Yu, Zhengyu Lv, Lixin Wang, Xiaomin Fang, Liurong Chen, Huanchun He, Kongwang |
description | The recently discovered porcine boca-like virus (Pbo-likeV) is a member of the Parvoviridae family, genus Bocavirus, and it is potentially associated with swine disease. Several studies have associated Pbo-likeV with postweaning multisystemic wasting syndrome in pigs, but the full spectrum of clinical disease and the epidemiology of Pbo-likeV infection remain unclear. The availability of rapid and reliable molecular diagnostics would aid future studies of this novel virus. Thus, we developed a sensitive and specific TaqMan-based real-time PCR assay to target the Pbo-likeV NP1 gene. The assay reproducibly detected 20 copies of a recombinant DNA plasmid containing the NP1 gene, with a dynamic range of six orders of magnitude (10(2)-10(7) copies). The assay did not cross-react with other animal viruses. Clinical evaluation found that Pbo-likeV was present in Chinese swine herds at a frequency of 44.2% (114/258). Higher infection rates were found in diseased pigs (56.1%, 101/180) compared with healthy pigs (16.7%, 13/78) (P < 0.05). Our assay for the diagnosis and quantification of Pbo-likeV was highly sensitive and specific, and should provide a reliable real-time tool for epidemiological and pathogenetic study of Pbo-likeV infection. |
doi_str_mv | 10.1186/1743-422X-8-357 |
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Several studies have associated Pbo-likeV with postweaning multisystemic wasting syndrome in pigs, but the full spectrum of clinical disease and the epidemiology of Pbo-likeV infection remain unclear. The availability of rapid and reliable molecular diagnostics would aid future studies of this novel virus. Thus, we developed a sensitive and specific TaqMan-based real-time PCR assay to target the Pbo-likeV NP1 gene. The assay reproducibly detected 20 copies of a recombinant DNA plasmid containing the NP1 gene, with a dynamic range of six orders of magnitude (10(2)-10(7) copies). The assay did not cross-react with other animal viruses. Clinical evaluation found that Pbo-likeV was present in Chinese swine herds at a frequency of 44.2% (114/258). Higher infection rates were found in diseased pigs (56.1%, 101/180) compared with healthy pigs (16.7%, 13/78) (P < 0.05). Our assay for the diagnosis and quantification of Pbo-likeV was highly sensitive and specific, and should provide a reliable real-time tool for epidemiological and pathogenetic study of Pbo-likeV infection.</description><identifier>ISSN: 1743-422X</identifier><identifier>EISSN: 1743-422X</identifier><identifier>DOI: 10.1186/1743-422X-8-357</identifier><identifier>PMID: 21771316</identifier><language>eng</language><publisher>England: BioMed Central Ltd</publisher><subject>Animals ; Bocavirus - isolation & purification ; Clinical Laboratory Techniques - methods ; Diseases ; DNA Primers - genetics ; DNA, Viral - genetics ; Genetic aspects ; Oligonucleotide Probes - chemistry ; Oligonucleotide Probes - genetics ; Parvoviridae Infections - diagnosis ; Parvoviridae Infections - veterinary ; Parvoviridae Infections - virology ; Parvoviruses ; Polymerase chain reaction ; Polymerase Chain Reaction - methods ; Reproducibility of Results ; Sensitivity and Specificity ; Short Report ; Swine ; Swine Diseases - diagnosis ; Swine Diseases - virology ; Viral Nonstructural Proteins - genetics ; Virology - methods</subject><ispartof>Virology journal, 2011-07, Vol.8 (1), p.357-357, Article 357</ispartof><rights>COPYRIGHT 2011 BioMed Central Ltd.</rights><rights>Copyright ©2011 Li et al; licensee BioMed Central Ltd. 2011 Li et al; licensee BioMed Central Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b618t-d08dc0f2e9850ba7aa79d7039f49e9f10cb22624db1d9d33f7c1ea9d0e94a4873</citedby><cites>FETCH-LOGICAL-b618t-d08dc0f2e9850ba7aa79d7039f49e9f10cb22624db1d9d33f7c1ea9d0e94a4873</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3152913/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3152913/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,27924,27925,37013,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21771316$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, Bin</creatorcontrib><creatorcontrib>Xiao, Shaobo</creatorcontrib><creatorcontrib>Ma, Junjie</creatorcontrib><creatorcontrib>Liu, Yanling</creatorcontrib><creatorcontrib>Mao, Li</creatorcontrib><creatorcontrib>Wen, Libin</creatorcontrib><creatorcontrib>Mao, Aihua</creatorcontrib><creatorcontrib>Zhang, Xuehan</creatorcontrib><creatorcontrib>Ni, Yanxiu</creatorcontrib><creatorcontrib>Guo, Rongli</creatorcontrib><creatorcontrib>Zhou, Junming</creatorcontrib><creatorcontrib>Yu, Zhengyu</creatorcontrib><creatorcontrib>Lv, Lixin</creatorcontrib><creatorcontrib>Wang, Xiaomin</creatorcontrib><creatorcontrib>Fang, Liurong</creatorcontrib><creatorcontrib>Chen, Huanchun</creatorcontrib><creatorcontrib>He, Kongwang</creatorcontrib><title>Development of a novel TaqMan-based real-time PCR assay for the detection of porcine boca-like virus (Pbo-likeV)</title><title>Virology journal</title><addtitle>Virol J</addtitle><description>The recently discovered porcine boca-like virus (Pbo-likeV) is a member of the Parvoviridae family, genus Bocavirus, and it is potentially associated with swine disease. Several studies have associated Pbo-likeV with postweaning multisystemic wasting syndrome in pigs, but the full spectrum of clinical disease and the epidemiology of Pbo-likeV infection remain unclear. The availability of rapid and reliable molecular diagnostics would aid future studies of this novel virus. Thus, we developed a sensitive and specific TaqMan-based real-time PCR assay to target the Pbo-likeV NP1 gene. The assay reproducibly detected 20 copies of a recombinant DNA plasmid containing the NP1 gene, with a dynamic range of six orders of magnitude (10(2)-10(7) copies). The assay did not cross-react with other animal viruses. Clinical evaluation found that Pbo-likeV was present in Chinese swine herds at a frequency of 44.2% (114/258). Higher infection rates were found in diseased pigs (56.1%, 101/180) compared with healthy pigs (16.7%, 13/78) (P < 0.05). Our assay for the diagnosis and quantification of Pbo-likeV was highly sensitive and specific, and should provide a reliable real-time tool for epidemiological and pathogenetic study of Pbo-likeV infection.</description><subject>Animals</subject><subject>Bocavirus - isolation & purification</subject><subject>Clinical Laboratory Techniques - methods</subject><subject>Diseases</subject><subject>DNA Primers - genetics</subject><subject>DNA, Viral - genetics</subject><subject>Genetic aspects</subject><subject>Oligonucleotide Probes - chemistry</subject><subject>Oligonucleotide Probes - genetics</subject><subject>Parvoviridae Infections - diagnosis</subject><subject>Parvoviridae Infections - veterinary</subject><subject>Parvoviridae Infections - virology</subject><subject>Parvoviruses</subject><subject>Polymerase chain reaction</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Short Report</subject><subject>Swine</subject><subject>Swine Diseases - diagnosis</subject><subject>Swine Diseases - virology</subject><subject>Viral Nonstructural Proteins - genetics</subject><subject>Virology - methods</subject><issn>1743-422X</issn><issn>1743-422X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNp1kstv1DAQxiMEoqVw5oYscYAe0vqRxPEFqSyvlYqoSkHcLD8mW5ck3trZFf3vcZqyaqQiH2yPv_nJ38xk2UuCjwipq2PCC5YXlP7K65yV_FG2v4s8vnfey57FeIUxoxUXT7M9SjgnjFT72foDbKH16w76AfkGKdT7FEAX6vqr6nOtIlgUQLX54DpAZ4tzpGJUN6jxAQ2XgCwMYAbn-zF77YNxPSDtjcpb9xvQ1oVNRG_PtL-9_zx8nj1pVBvhxd1-kP349PFi8SU__fZ5uTg5zXVF6iG3uLYGNxREXWKtuFJcWI6ZaAoBoiHYaEorWlhNrLCMNdwQUMJiEIUqas4OsuXEtV5dyXVwnQo30isnbwM-rKQKgzMtSNA115rQRvOyqKHQRFSJqcuyqjBjOLHeTaz1RndgTapVUO0MOn_p3aVc-a1kpKSCsAR4PwG08_8BzF-M7-TYPTl2T9Yy9TZB3tz9IvjrDcRBdi4aaFvVg99EWdcMYy5EmZSvJ-VKJXuub3yCmlEtT2hV0jQGYnR19IAqLQudM76HxqX4LOFwlpA0A_wZVmoTo1x-P59rjyetCT7GAM3OLMFyHN0H7L26X-Sd_t-ssr8v9-hL</recordid><startdate>20110719</startdate><enddate>20110719</enddate><creator>Li, Bin</creator><creator>Xiao, Shaobo</creator><creator>Ma, Junjie</creator><creator>Liu, Yanling</creator><creator>Mao, Li</creator><creator>Wen, Libin</creator><creator>Mao, Aihua</creator><creator>Zhang, Xuehan</creator><creator>Ni, Yanxiu</creator><creator>Guo, Rongli</creator><creator>Zhou, Junming</creator><creator>Yu, Zhengyu</creator><creator>Lv, Lixin</creator><creator>Wang, Xiaomin</creator><creator>Fang, Liurong</creator><creator>Chen, Huanchun</creator><creator>He, Kongwang</creator><general>BioMed Central Ltd</general><general>BioMed Central</general><general>BMC</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>ISR</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20110719</creationdate><title>Development of a novel TaqMan-based real-time PCR assay for the detection of porcine boca-like virus (Pbo-likeV)</title><author>Li, Bin ; Xiao, Shaobo ; Ma, Junjie ; Liu, Yanling ; Mao, Li ; Wen, Libin ; Mao, Aihua ; Zhang, Xuehan ; Ni, Yanxiu ; Guo, Rongli ; Zhou, Junming ; Yu, Zhengyu ; Lv, Lixin ; Wang, Xiaomin ; Fang, Liurong ; Chen, Huanchun ; He, Kongwang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b618t-d08dc0f2e9850ba7aa79d7039f49e9f10cb22624db1d9d33f7c1ea9d0e94a4873</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Bocavirus - isolation & purification</topic><topic>Clinical Laboratory Techniques - methods</topic><topic>Diseases</topic><topic>DNA Primers - genetics</topic><topic>DNA, Viral - genetics</topic><topic>Genetic aspects</topic><topic>Oligonucleotide Probes - chemistry</topic><topic>Oligonucleotide Probes - genetics</topic><topic>Parvoviridae Infections - diagnosis</topic><topic>Parvoviridae Infections - veterinary</topic><topic>Parvoviridae Infections - virology</topic><topic>Parvoviruses</topic><topic>Polymerase chain reaction</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Short Report</topic><topic>Swine</topic><topic>Swine Diseases - diagnosis</topic><topic>Swine Diseases - virology</topic><topic>Viral Nonstructural Proteins - genetics</topic><topic>Virology - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, Bin</creatorcontrib><creatorcontrib>Xiao, Shaobo</creatorcontrib><creatorcontrib>Ma, Junjie</creatorcontrib><creatorcontrib>Liu, Yanling</creatorcontrib><creatorcontrib>Mao, Li</creatorcontrib><creatorcontrib>Wen, Libin</creatorcontrib><creatorcontrib>Mao, Aihua</creatorcontrib><creatorcontrib>Zhang, Xuehan</creatorcontrib><creatorcontrib>Ni, Yanxiu</creatorcontrib><creatorcontrib>Guo, Rongli</creatorcontrib><creatorcontrib>Zhou, Junming</creatorcontrib><creatorcontrib>Yu, Zhengyu</creatorcontrib><creatorcontrib>Lv, Lixin</creatorcontrib><creatorcontrib>Wang, Xiaomin</creatorcontrib><creatorcontrib>Fang, Liurong</creatorcontrib><creatorcontrib>Chen, Huanchun</creatorcontrib><creatorcontrib>He, Kongwang</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Gale In Context: Science</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>Directory of Open Access Journals</collection><jtitle>Virology journal</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, Bin</au><au>Xiao, Shaobo</au><au>Ma, Junjie</au><au>Liu, Yanling</au><au>Mao, Li</au><au>Wen, Libin</au><au>Mao, Aihua</au><au>Zhang, Xuehan</au><au>Ni, Yanxiu</au><au>Guo, Rongli</au><au>Zhou, Junming</au><au>Yu, Zhengyu</au><au>Lv, Lixin</au><au>Wang, Xiaomin</au><au>Fang, Liurong</au><au>Chen, Huanchun</au><au>He, Kongwang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Development of a novel TaqMan-based real-time PCR assay for the detection of porcine boca-like virus (Pbo-likeV)</atitle><jtitle>Virology journal</jtitle><addtitle>Virol J</addtitle><date>2011-07-19</date><risdate>2011</risdate><volume>8</volume><issue>1</issue><spage>357</spage><epage>357</epage><pages>357-357</pages><artnum>357</artnum><issn>1743-422X</issn><eissn>1743-422X</eissn><abstract>The recently discovered porcine boca-like virus (Pbo-likeV) is a member of the Parvoviridae family, genus Bocavirus, and it is potentially associated with swine disease. Several studies have associated Pbo-likeV with postweaning multisystemic wasting syndrome in pigs, but the full spectrum of clinical disease and the epidemiology of Pbo-likeV infection remain unclear. The availability of rapid and reliable molecular diagnostics would aid future studies of this novel virus. Thus, we developed a sensitive and specific TaqMan-based real-time PCR assay to target the Pbo-likeV NP1 gene. The assay reproducibly detected 20 copies of a recombinant DNA plasmid containing the NP1 gene, with a dynamic range of six orders of magnitude (10(2)-10(7) copies). The assay did not cross-react with other animal viruses. Clinical evaluation found that Pbo-likeV was present in Chinese swine herds at a frequency of 44.2% (114/258). Higher infection rates were found in diseased pigs (56.1%, 101/180) compared with healthy pigs (16.7%, 13/78) (P < 0.05). Our assay for the diagnosis and quantification of Pbo-likeV was highly sensitive and specific, and should provide a reliable real-time tool for epidemiological and pathogenetic study of Pbo-likeV infection.</abstract><cop>England</cop><pub>BioMed Central Ltd</pub><pmid>21771316</pmid><doi>10.1186/1743-422X-8-357</doi><oa>free_for_read</oa></addata></record> |
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subjects | Animals Bocavirus - isolation & purification Clinical Laboratory Techniques - methods Diseases DNA Primers - genetics DNA, Viral - genetics Genetic aspects Oligonucleotide Probes - chemistry Oligonucleotide Probes - genetics Parvoviridae Infections - diagnosis Parvoviridae Infections - veterinary Parvoviridae Infections - virology Parvoviruses Polymerase chain reaction Polymerase Chain Reaction - methods Reproducibility of Results Sensitivity and Specificity Short Report Swine Swine Diseases - diagnosis Swine Diseases - virology Viral Nonstructural Proteins - genetics Virology - methods |
title | Development of a novel TaqMan-based real-time PCR assay for the detection of porcine boca-like virus (Pbo-likeV) |
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