Identification of the severe acute respiratory syndrome coronavirus by simultaneous multigene DNA sequencing.
Identifieur interne : 002C69 ( PubMed/Corpus ); précédent : 002C68; suivant : 002C70Identification of the severe acute respiratory syndrome coronavirus by simultaneous multigene DNA sequencing.
Auteurs : T. Vinayagamoorthy ; Kirk Mulatz ; Roger HodkinsonSource :
- Journal of clinical microbiology [ 0095-1137 ] ; 2004.
English descriptors
- KwdEn :
- MESH :
- genetics : SARS Virus.
- isolation & purification : SARS Virus.
- Plasmids, Polymerase Chain Reaction, Sequence Analysis, DNA, Transformation, Genetic.
Abstract
The recent severe acute respiratory syndrome (SARS) outbreak resulted in calls for an accurate diagnostic test that can be used not only for routine testing but also for generating nucleotide sequences to monitor the epidemic. Although the identity of the SARS coronavirus (SARS-CoV) genome was confirmed by DNA sequencing, it is impractical to sequence the entire 29-kb SARS-CoV genome on a routine basis. Therefore, alternative assay methods such as the enzyme-linked immunosorbent assay and PCR have been pursued for routine testing, primarily to resolve probable cases. We report here a modification of standard DNA sequencing technology for accurate identification of SARS-CoV in routine testing. Instead of requiring the sequencing of the whole SARS-CoV genome, our modification enables the simultaneous sequencing of three regions of the SARS-CoV genome, the spike protein-encoding gene (35 nucleotides), gene M (43 nucleotides), and gene N (45 nucleotides), in a single electropherogram. Comparing these nucleotide sequences to DNA databank entries (National Institutes of Health) conclusively identified them as SARS-CoV sequences.
DOI: 10.1128/JCM.42.7.3291-3294.2004
PubMed: 15243096
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pubmed:15243096Le document en format XML
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Vinayagamoorthy</name><affiliation><nlm:affiliation>Bio-ID Diagnostic Inc., No. 1, 410 Downey Rd., LFK Biotechnology Complex, Saskatoon, Saskatchewan, Canada S7N 4N1. moorthy@bio-id-diagnostic.com</nlm:affiliation></affiliation></author><author><name sortKey="Mulatz, Kirk" sort="Mulatz, Kirk" uniqKey="Mulatz K" first="Kirk" last="Mulatz">Kirk Mulatz</name></author><author><name sortKey="Hodkinson, Roger" sort="Hodkinson, Roger" uniqKey="Hodkinson R" first="Roger" last="Hodkinson">Roger Hodkinson</name></author></analytic><series><title level="j">Journal of clinical microbiology</title><idno type="ISSN">0095-1137</idno><imprint><date when="2004" type="published">2004</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Plasmids</term><term>Polymerase Chain Reaction</term><term>SARS Virus (genetics)</term><term>SARS Virus (isolation & purification)</term><term>Sequence Analysis, DNA</term><term>Transformation, Genetic</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" qualifier="isolation & purification" xml:lang="en"><term>SARS Virus</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Plasmids</term><term>Polymerase Chain Reaction</term><term>Sequence Analysis, DNA</term><term>Transformation, Genetic</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The recent severe acute respiratory syndrome (SARS) outbreak resulted in calls for an accurate diagnostic test that can be used not only for routine testing but also for generating nucleotide sequences to monitor the epidemic. Although the identity of the SARS coronavirus (SARS-CoV) genome was confirmed by DNA sequencing, it is impractical to sequence the entire 29-kb SARS-CoV genome on a routine basis. Therefore, alternative assay methods such as the enzyme-linked immunosorbent assay and PCR have been pursued for routine testing, primarily to resolve probable cases. We report here a modification of standard DNA sequencing technology for accurate identification of SARS-CoV in routine testing. Instead of requiring the sequencing of the whole SARS-CoV genome, our modification enables the simultaneous sequencing of three regions of the SARS-CoV genome, the spike protein-encoding gene (35 nucleotides), gene M (43 nucleotides), and gene N (45 nucleotides), in a single electropherogram. Comparing these nucleotide sequences to DNA databank entries (National Institutes of Health) conclusively identified them as SARS-CoV sequences.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">15243096</PMID><DateCompleted><Year>2004</Year><Month>08</Month><Day>19</Day></DateCompleted><DateRevised><Year>2018</Year><Month>11</Month><Day>13</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0095-1137</ISSN><JournalIssue CitedMedium="Print"><Volume>42</Volume><Issue>7</Issue><PubDate><Year>2004</Year><Month>Jul</Month></PubDate></JournalIssue><Title>Journal of clinical microbiology</Title><ISOAbbreviation>J. Clin. 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Instead of requiring the sequencing of the whole SARS-CoV genome, our modification enables the simultaneous sequencing of three regions of the SARS-CoV genome, the spike protein-encoding gene (35 nucleotides), gene M (43 nucleotides), and gene N (45 nucleotides), in a single electropherogram. 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