Identification and subtyping of avian influenza viruses by reverse transcription-PCR
Identifieur interne : 000C00 ( Istex/Corpus ); précédent : 000B99; suivant : 000C01Identification and subtyping of avian influenza viruses by reverse transcription-PCR
Auteurs : Ming-Shiuh Lee ; Poa-Chun Chang ; Jui-Hung Shien ; Ming-Chu Cheng ; Happy K. ShiehSource :
- Journal of Virological Methods [ 0166-0934 ] ; 2001.
English descriptors
- KwdEn :
- Teeft :
- A6ian 6irus, Accession numbers, Avian, Avian virus, Avian viruses, Blast search, Clin, Embryonated eggs, Equine origins, Genbank, Hemagglutinin, Homogenate, Hong kong, Microbiol, Nucleotide sequence, Nucleotide sequences, Organ homogenates, Primer, Primer design, Reference center, Reference strains, Sequence analysis, Sequence comparison, Subtype, Subtypes, Subtyping, Veterinary medicine, Virol, Virological, Virological methods, Virus.
Abstract
Abstract: Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1–H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate, only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure, and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates.
Url:
DOI: 10.1016/S0166-0934(01)00301-9
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ISTEX:5D2576E3791CB87E4885E44FE07686427B2B838BLe document en format XML
<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title xml:lang="en">Identification and subtyping of avian influenza viruses by reverse transcription-PCR</title><author><name sortKey="Lee, Ming Shiuh" sort="Lee, Ming Shiuh" uniqKey="Lee M" first="Ming-Shiuh" last="Lee">Ming-Shiuh Lee</name><affiliation><mods:affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</mods:affiliation></affiliation></author><author><name sortKey="Chang, Poa Chun" sort="Chang, Poa Chun" uniqKey="Chang P" first="Poa-Chun" last="Chang">Poa-Chun Chang</name><affiliation><mods:affiliation>Institute of Veterinary Microbiology, National Chung Hsing University, Taichung, Taiwan</mods:affiliation></affiliation></author><author><name sortKey="Shien, Jui Hung" sort="Shien, Jui Hung" uniqKey="Shien J" first="Jui-Hung" last="Shien">Jui-Hung Shien</name><affiliation><mods:affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</mods:affiliation></affiliation></author><author><name sortKey="Cheng, Ming Chu" sort="Cheng, Ming Chu" uniqKey="Cheng M" first="Ming-Chu" last="Cheng">Ming-Chu Cheng</name><affiliation><mods:affiliation>National Institute of Animal Health, Council of Agriculture, Taipei, Taiwan</mods:affiliation></affiliation></author><author><name sortKey="Shieh, Happy K" sort="Shieh, Happy K" uniqKey="Shieh H" first="Happy K" last="Shieh">Happy K. 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Shieh</name><affiliation><mods:affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="ISSN">0166-0934</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2001">2001</date><biblScope unit="volume">97</biblScope><biblScope unit="issue">1–2</biblScope><biblScope unit="page" from="13">13</biblScope><biblScope unit="page" to="22">22</biblScope></imprint><idno type="ISSN">0166-0934</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0166-0934</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Avian influenza virus</term><term>Hemagglutinin</term><term>Reverse transcription-polymerase chain reaction (RT-PCR)</term><term>Subtype</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>A6ian 6irus</term><term>Accession numbers</term><term>Avian</term><term>Avian virus</term><term>Avian viruses</term><term>Blast search</term><term>Clin</term><term>Embryonated eggs</term><term>Equine origins</term><term>Genbank</term><term>Hemagglutinin</term><term>Homogenate</term><term>Hong kong</term><term>Microbiol</term><term>Nucleotide sequence</term><term>Nucleotide sequences</term><term>Organ homogenates</term><term>Primer</term><term>Primer design</term><term>Reference center</term><term>Reference strains</term><term>Sequence analysis</term><term>Sequence comparison</term><term>Subtype</term><term>Subtypes</term><term>Subtyping</term><term>Veterinary medicine</term><term>Virol</term><term>Virological</term><term>Virological methods</term><term>Virus</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1–H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate, only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure, and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates.</div></front></TEI><istex><corpusName>elsevier</corpusName><keywords><teeft><json:string>avian</json:string><json:string>primer</json:string><json:string>avian viruses</json:string><json:string>subtyping</json:string><json:string>avian virus</json:string><json:string>subtypes</json:string><json:string>virological</json:string><json:string>genbank</json:string><json:string>virological methods</json:string><json:string>organ homogenates</json:string><json:string>hemagglutinin</json:string><json:string>microbiol</json:string><json:string>virol</json:string><json:string>clin</json:string><json:string>reference strains</json:string><json:string>primer design</json:string><json:string>sequence comparison</json:string><json:string>sequence analysis</json:string><json:string>hong kong</json:string><json:string>virus</json:string><json:string>nucleotide sequences</json:string><json:string>homogenate</json:string><json:string>reference center</json:string><json:string>equine origins</json:string><json:string>nucleotide sequence</json:string><json:string>embryonated eggs</json:string><json:string>a6ian 6irus</json:string><json:string>blast search</json:string><json:string>accession numbers</json:string><json:string>veterinary medicine</json:string><json:string>subtype</json:string></teeft></keywords><author><json:item><name>Ming-Shiuh Lee</name><affiliations><json:string>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</json:string></affiliations></json:item><json:item><name>Poa-Chun Chang</name><affiliations><json:string>Institute of Veterinary Microbiology, National Chung Hsing University, Taichung, Taiwan</json:string></affiliations></json:item><json:item><name>Jui-Hung Shien</name><affiliations><json:string>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</json:string></affiliations></json:item><json:item><name>Ming-Chu Cheng</name><affiliations><json:string>National Institute of Animal Health, Council of Agriculture, Taipei, Taiwan</json:string></affiliations></json:item><json:item><name>Happy K Shieh</name><affiliations><json:string>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>Avian influenza virus</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Hemagglutinin</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Subtype</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>Reverse transcription-polymerase chain reaction (RT-PCR)</value></json:item></subject><arkIstex>ark:/67375/6H6-JZ2VDGG3-Z</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>Full-length article</json:string></originalGenre><abstract>Abstract: Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1–H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate, only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure, and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates.</abstract><qualityIndicators><score>8.332</score><pdfWordCount>4184</pdfWordCount><pdfCharCount>26945</pdfCharCount><pdfVersion>1.2</pdfVersion><pdfPageCount>10</pdfPageCount><pdfPageSize>536 x 674 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>179</abstractWordCount><abstractCharCount>1147</abstractCharCount><keywordCount>4</keywordCount></qualityIndicators><title>Identification and subtyping of avian influenza viruses by reverse transcription-PCR</title><pmid><json:string>11483213</json:string></pmid><pii><json:string>S0166-0934(01)00301-9</json:string></pii><genre><json:string>research-article</json:string></genre><host><title>Journal of Virological Methods</title><language><json:string>unknown</json:string></language><publicationDate>2001</publicationDate><issn><json:string>0166-0934</json:string></issn><pii><json:string>S0166-0934(00)X0079-1</json:string></pii><volume>97</volume><issue>1–2</issue><pages><first>13</first><last>22</last></pages><genre><json:string>journal</json:string></genre></host><namedEntities><unitex><date><json:string>1997</json:string><json:string>2001</json:string><json:string>1999</json:string><json:string>1989</json:string></date><geogName></geogName><orgName><json:string>NCBI</json:string><json:string>Taiwan Received</json:string><json:string>DNASTAR Inc., Madison</json:string><json:string>National Center of Biotechnology Information</json:string><json:string>Hokkaido University</json:string><json:string>Turkey/Ontario</json:string><json:string>National Institute of Animal Health, Council of Agriculture, Taipei, Taiwan</json:string><json:string>Taiwan, Republic of China</json:string><json:string>Institute of Veterinary Microbiology, National Chung Hsing Uni</json:string><json:string>Jude Children’s Research Hospital, Memphis, TN</json:string><json:string>Health</json:string><json:string>Turkey/Wisconsin</json:string><json:string>QIAGEN Inc., Valencia</json:string></orgName><orgName_funder></orgName_funder><orgName_provider></orgName_provider><persName><json:string>Hong Kong</json:string><json:string>H. 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Lee et al.</json:string><json:string>Senne et al., 1996</json:string><json:string>Wiley et al., 1981</json:string><json:string>Alexander, 1995</json:string><json:string>Cauthen et al., 2000</json:string><json:string>Beard 1989a,b</json:string><json:string>Altschul et al., 1990</json:string><json:string>Air (1981)</json:string><json:string>Webster et al., 1997</json:string><json:string>Suarez et al., 1998</json:string><json:string>Claas et al., 1993</json:string><json:string>Rohm et al., 1996</json:string><json:string>Shu et al., 1993</json:string><json:string>Altmuller et al., 1989</json:string><json:string>Claas et al., 1998</json:string><json:string>Yuen et al., 1998</json:string><json:string>Wright et al., 1995</json:string><json:string>Zhou et al., 1996</json:string><json:string>Lee et al.</json:string><json:string>Subbarao et al., 1998</json:string><json:string>Lamb and Krug, 1996</json:string><json:string>Hoffmann et al., 2000</json:string><json:string>Cherian et al., 1994</json:string><json:string>Zou, 1997</json:string><json:string>Beard, 1989b</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/6H6-JZ2VDGG3-Z</json:string></ark><categories><wos><json:string>1 - science</json:string><json:string>2 - virology</json:string><json:string>2 - biotechnology & applied microbiology</json:string><json:string>2 - biochemical research methods</json:string></wos><scienceMetrix><json:string>1 - health sciences</json:string><json:string>2 - biomedical research</json:string><json:string>3 - virology</json:string></scienceMetrix><scopus><json:string>1 - Life Sciences</json:string><json:string>2 - Immunology and Microbiology</json:string><json:string>3 - Virology</json:string></scopus><inist><json:string>1 - sciences appliquees, technologies et medecines</json:string><json:string>2 - sciences biologiques et medicales</json:string><json:string>3 - sciences medicales</json:string></inist></categories><publicationDate>2001</publicationDate><copyrightDate>2001</copyrightDate><doi><json:string>10.1016/S0166-0934(01)00301-9</json:string></doi><id>5D2576E3791CB87E4885E44FE07686427B2B838B</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-JZ2VDGG3-Z/fulltext.pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-JZ2VDGG3-Z/bundle.zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/ark:/67375/6H6-JZ2VDGG3-Z/fulltext.tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Identification and subtyping of avian influenza viruses by reverse transcription-PCR</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher scheme="https://scientific-publisher.data.istex.fr">ELSEVIER</publisher><availability><licence><p>©2001 Elsevier Science B.V.</p></licence><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</p></availability><date>2001</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note><note type="content">Fig. 1: HA-subtyping of avian influenza viruses by RT-PCR. Each panel is the subtyping result of a single reference strain, of which the subtype is shown on the left top of each panel. The expected sizes of RT-PCR products are shown in parentheses. Lane M, size markers (100 bp ladder, PRO-tech, Taipei, Taiwan); lanes 1–15, RT-PCR products after amplification with primers specific to H1–H15 subtypes; lane C, negative control.</note><note type="content">Fig. 2: Identification and subtyping of avian influenza viruses in organ homogenates. Lane M, size markers (100 bp ladder, PRO-tech, Taipei, Taiwan); lanes 1–15, RT-PCR products obtained by using primers specific to H1–H15 subtypes; lane 16, the RT-PCR product obtained by using primers specific to the NP gene of the virus. The sizes of RT-PCR products, 302 bp for H6 and 330 bp for NP gene, are indicated by arrows.</note><note type="content">Table 1: Reference strains and strains sequenced in this study</note><note type="content">Table 2: Primers used for HA-subtyping of avian influenza viruses by RT-PCR</note><note type="content">Table 3: Comparison of the result of HA-subtyping by RT-PCR and by HI test</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Identification and subtyping of avian influenza viruses by reverse transcription-PCR</title><author xml:id="author-0000"><persName><forename type="first">Ming-Shiuh</forename><surname>Lee</surname></persName><affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</affiliation></author><author xml:id="author-0001"><persName><forename type="first">Poa-Chun</forename><surname>Chang</surname></persName><affiliation>Institute of Veterinary Microbiology, National Chung Hsing University, Taichung, Taiwan</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Jui-Hung</forename><surname>Shien</surname></persName><affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</affiliation></author><author xml:id="author-0003"><persName><forename type="first">Ming-Chu</forename><surname>Cheng</surname></persName><affiliation>National Institute of Animal Health, Council of Agriculture, Taipei, Taiwan</affiliation></author><author xml:id="author-0004"><persName><forename type="first">Happy K</forename><surname>Shieh</surname></persName><note type="correspondence"><p>Corresponding author. Tel.: +886-4-22860196; Fax: +886-4-22872392</p></note><affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</affiliation></author><idno type="istex">5D2576E3791CB87E4885E44FE07686427B2B838B</idno><idno type="ark">ark:/67375/6H6-JZ2VDGG3-Z</idno><idno type="DOI">10.1016/S0166-0934(01)00301-9</idno><idno type="PII">S0166-0934(01)00301-9</idno></analytic><monogr><title level="j">Journal of Virological Methods</title><title level="j" type="abbrev">VIRMET</title><idno type="pISSN">0166-0934</idno><idno type="PII">S0166-0934(00)X0079-1</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="2001"></date><biblScope unit="volume">97</biblScope><biblScope unit="issue">1–2</biblScope><biblScope unit="page" from="13">13</biblScope><biblScope unit="page" to="22">22</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2001</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Abstract: Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1–H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate, only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure, and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates.</p></abstract><textClass xml:lang="en"><keywords scheme="keyword"><list><head>Keywords</head><item><term>Avian influenza virus</term></item><item><term>Hemagglutinin</term></item><item><term>Subtype</term></item><item><term>Reverse transcription-polymerase chain reaction (RT-PCR)</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2001-03-05">Modified</change><change when="2001">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-JZ2VDGG3-Z/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: ce:floats; body; tail"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"><istex:entity SYSTEM="gr1y1" NDATA="IMAGE" name="gr1y1"></istex:entity><istex:entity SYSTEM="gr1y2" NDATA="IMAGE" name="gr1y2"></istex:entity><istex:entity SYSTEM="gr2" NDATA="IMAGE" name="gr2"></istex:entity></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla" xml:lang="en"><item-info><jid>VIRMET</jid><aid>2924</aid><ce:pii>S0166-0934(01)00301-9</ce:pii><ce:doi>10.1016/S0166-0934(01)00301-9</ce:doi><ce:copyright type="full-transfer" year="2001">Elsevier Science B.V.</ce:copyright></item-info><head><ce:title>Identification and subtyping of avian influenza viruses by reverse transcription-PCR</ce:title><ce:author-group><ce:author><ce:given-name>Ming-Shiuh</ce:given-name><ce:surname>Lee</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Poa-Chun</ce:given-name><ce:surname>Chang</ce:surname><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Jui-Hung</ce:given-name><ce:surname>Shien</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Ming-Chu</ce:given-name><ce:surname>Cheng</ce:surname><ce:cross-ref refid="AFF2"><ce:sup>b</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Happy K</ce:given-name><ce:surname>Shieh</ce:surname><ce:cross-ref refid="AFF1"><ce:sup>a</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF3"><ce:sup>c</ce:sup></ce:cross-ref><ce:cross-ref refid="CORR1">*</ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>National Institute of Animal Health, Council of Agriculture, Taipei, Taiwan</ce:textfn></ce:affiliation><ce:affiliation id="AFF3"><ce:label>c</ce:label><ce:textfn>Institute of Veterinary Microbiology, National Chung Hsing University, Taichung, Taiwan</ce:textfn></ce:affiliation><ce:correspondence id="CORR1"><ce:label>*</ce:label><ce:text>Corresponding author. Tel.: +886-4-22860196; Fax: +886-4-22872392</ce:text></ce:correspondence></ce:author-group><ce:date-received day="6" month="11" year="2000"></ce:date-received><ce:date-revised day="5" month="3" year="2001"></ce:date-revised><ce:date-accepted day="7" month="3" year="2001"></ce:date-accepted><ce:abstract><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para>Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1–H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate, only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure, and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates.</ce:simple-para></ce:abstract-sec></ce:abstract><ce:keywords class="keyword"><ce:section-title>Keywords</ce:section-title><ce:keyword><ce:text>Avian influenza virus</ce:text></ce:keyword><ce:keyword><ce:text>Hemagglutinin</ce:text></ce:keyword><ce:keyword><ce:text>Subtype</ce:text></ce:keyword><ce:keyword><ce:text>Reverse transcription-polymerase chain reaction (RT-PCR)</ce:text></ce:keyword></ce:keywords></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Identification and subtyping of avian influenza viruses by reverse transcription-PCR</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Identification and subtyping of avian influenza viruses by reverse transcription-PCR</title></titleInfo><name type="personal"><namePart type="given">Ming-Shiuh</namePart><namePart type="family">Lee</namePart><affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Poa-Chun</namePart><namePart type="family">Chang</namePart><affiliation>Institute of Veterinary Microbiology, National Chung Hsing University, Taichung, Taiwan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Jui-Hung</namePart><namePart type="family">Shien</namePart><affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ming-Chu</namePart><namePart type="family">Cheng</namePart><affiliation>National Institute of Animal Health, Council of Agriculture, Taipei, Taiwan</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Happy K</namePart><namePart type="family">Shieh</namePart><affiliation>Department of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan</affiliation><description>Corresponding author. Tel.: +886-4-22860196; Fax: +886-4-22872392</description><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">2001</dateIssued><dateModified encoding="w3cdtf">2001-03-05</dateModified><copyrightDate encoding="w3cdtf">2001</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: Avian influenza viruses have 15 different hemagglutinin (HA) subtypes (H1–H15). We report a procedure for the identification and HA-subtyping of avian influenza virus by reverse transcription-PCR (RT-PCR). The avian influenza virus is identified by RT-PCR using a set of primers specific to the nucleoprotein (NP) gene of avian influenza virus. The HA-subtypes of avian influenza virus were determined by running simultaneously 15 RT-PCR reactions, each using a set of primers specific to one HA-subtype. For a single virus strain or isolate, only one of the 15 RT-PCR reactions will give a product of expected size, and thus the HA-subtype of the virus is determined. The result of HA-subtyping was then confirmed by sequence analysis of the PCR product. A total of 80 strains or isolates of avian influenza viruses were subtyped by this RT-PCR procedure, and the result of RT-PCR gave an excellent (100%) correlation with the result of the conventional serological method. The RT-PCR procedure we developed is rapid and sensitive, and could be used for the identification and HA-subtyping of avian influenza virus in organ homogenates.</abstract><note type="content">Fig. 1: HA-subtyping of avian influenza viruses by RT-PCR. Each panel is the subtyping result of a single reference strain, of which the subtype is shown on the left top of each panel. The expected sizes of RT-PCR products are shown in parentheses. Lane M, size markers (100 bp ladder, PRO-tech, Taipei, Taiwan); lanes 1–15, RT-PCR products after amplification with primers specific to H1–H15 subtypes; lane C, negative control.</note><note type="content">Fig. 2: Identification and subtyping of avian influenza viruses in organ homogenates. Lane M, size markers (100 bp ladder, PRO-tech, Taipei, Taiwan); lanes 1–15, RT-PCR products obtained by using primers specific to H1–H15 subtypes; lane 16, the RT-PCR product obtained by using primers specific to the NP gene of the virus. The sizes of RT-PCR products, 302 bp for H6 and 330 bp for NP gene, are indicated by arrows.</note><note type="content">Table 1: Reference strains and strains sequenced in this study</note><note type="content">Table 2: Primers used for HA-subtyping of avian influenza viruses by RT-PCR</note><note type="content">Table 3: Comparison of the result of HA-subtyping by RT-PCR and by HI test</note><subject lang="en"><genre>Keywords</genre><topic>Avian influenza virus</topic><topic>Hemagglutinin</topic><topic>Subtype</topic><topic>Reverse transcription-polymerase chain reaction (RT-PCR)</topic></subject><relatedItem type="host"><titleInfo><title>Journal of Virological Methods</title></titleInfo><titleInfo type="abbreviated"><title>VIRMET</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">2001</dateIssued></originInfo><identifier type="ISSN">0166-0934</identifier><identifier type="PII">S0166-0934(00)X0079-1</identifier><part><date>2001</date><detail type="volume"><number>97</number><caption>vol.</caption></detail><detail type="issue"><number>1–2</number><caption>no.</caption></detail><extent unit="issue-pages"><start>1</start><end>179</end></extent><extent unit="pages"><start>13</start><end>22</end></extent></part></relatedItem><identifier type="istex">5D2576E3791CB87E4885E44FE07686427B2B838B</identifier><identifier type="ark">ark:/67375/6H6-JZ2VDGG3-Z</identifier><identifier type="DOI">10.1016/S0166-0934(01)00301-9</identifier><identifier type="PII">S0166-0934(01)00301-9</identifier><accessCondition type="use and reproduction" contentType="copyright">©2001 Elsevier Science B.V.</accessCondition><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource><recordOrigin>Elsevier Science B.V., ©2001</recordOrigin></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-JZ2VDGG3-Z/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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