Direct detection of Staphylococcus aureus mRNA using a flow through microarray.
Identifieur interne : 002355 ( PubMed/Curation ); précédent : 002354; suivant : 002356Direct detection of Staphylococcus aureus mRNA using a flow through microarray.
Auteurs : R M Anthony [Pays-Bas] ; A R J. Schuitema ; L. Oskam ; P R KlatserSource :
- Journal of microbiological methods [ 0167-7012 ] ; 2005.
Descripteurs français
- KwdFr :
- ARN bactérien (), ARN bactérien (génétique), ARN bactérien (isolement et purification), ARN messager (), ARN messager (génétique), ARN messager (isolement et purification), ARN ribosomique 23S (), ARN ribosomique 23S (génétique), Humains, Staphylococcus aureus (génétique), Séquençage par oligonucléotides en batterie ().
- MESH :
- génétique : ARN bactérien, ARN messager, ARN ribosomique 23S, Staphylococcus aureus.
- isolement et purification : ARN bactérien, ARN messager.
- ARN bactérien, ARN messager, ARN ribosomique 23S, Humains, Séquençage par oligonucléotides en batterie.
English descriptors
- KwdEn :
- Humans, Oligonucleotide Array Sequence Analysis (methods), RNA, Bacterial (chemistry), RNA, Bacterial (genetics), RNA, Bacterial (isolation & purification), RNA, Messenger (chemistry), RNA, Messenger (genetics), RNA, Messenger (isolation & purification), RNA, Ribosomal, 23S (chemistry), RNA, Ribosomal, 23S (genetics), Staphylococcus aureus (genetics).
- MESH :
- chemical , chemistry : RNA, Bacterial, RNA, Messenger, RNA, Ribosomal, 23S.
- chemical , genetics : RNA, Bacterial, RNA, Messenger, RNA, Ribosomal, 23S.
- chemical , isolation & purification : RNA, Bacterial, RNA, Messenger.
- genetics : Staphylococcus aureus.
- methods : Oligonucleotide Array Sequence Analysis.
- Humans.
Abstract
The direct detection of mRNAs from bacterial cultures on a DNA array without amplification and labelling would greatly extend the range of applications suitable for microarray analysis. Here we describe the direct detection of 23S rRNA and seven mRNA species from total Staphylococcus aureus RNA prepared using commercially available RNA purification columns followed by fluorescent detection on a flow through microarray. RNA hybridisation was detected using paired secondary labelled probes directly 5' and 3' to immobilised 60 mers. In this way, we were able to detect the effect of 30-min exposure to antimicrobials on mRNA levels within 3 h after column purification of total RNA without the need for enzymatic manipulation. Specifically the expression of mecA was confirmed in a highly resistant strain and induction of katA and ile-tRNA synthetase genes after exposure to mupirocin could be detected.
DOI: 10.1016/j.mimet.2004.08.009
PubMed: 15567224
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Oskam</name></author><author><name sortKey="Klatser, P R" sort="Klatser, P R" uniqKey="Klatser P" first="P R" last="Klatser">P R Klatser</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:15567224</idno><idno type="pmid">15567224</idno><idno type="doi">10.1016/j.mimet.2004.08.009</idno><idno type="wicri:Area/PubMed/Corpus">002355</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002355</idno><idno type="wicri:Area/PubMed/Curation">002355</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002355</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Direct detection of Staphylococcus aureus mRNA using a flow through microarray.</title><author><name sortKey="Anthony, R M" sort="Anthony, R M" uniqKey="Anthony R" first="R M" last="Anthony">R M Anthony</name><affiliation wicri:level="1"><nlm:affiliation>KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam, The Netherlands. r.anthony@kit.nl</nlm:affiliation><country xml:lang="fr">Pays-Bas</country><wicri:regionArea>KIT Biomedical Research, Meibergdreef 39, 1105 AZ Amsterdam</wicri:regionArea></affiliation></author><author><name sortKey="Schuitema, A R J" sort="Schuitema, A R J" uniqKey="Schuitema A" first="A R J" last="Schuitema">A R J. 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Here we describe the direct detection of 23S rRNA and seven mRNA species from total Staphylococcus aureus RNA prepared using commercially available RNA purification columns followed by fluorescent detection on a flow through microarray. RNA hybridisation was detected using paired secondary labelled probes directly 5' and 3' to immobilised 60 mers. In this way, we were able to detect the effect of 30-min exposure to antimicrobials on mRNA levels within 3 h after column purification of total RNA without the need for enzymatic manipulation. Specifically the expression of mecA was confirmed in a highly resistant strain and induction of katA and ile-tRNA synthetase genes after exposure to mupirocin could be detected.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">15567224</PMID><DateCompleted><Year>2005</Year><Month>03</Month><Day>10</Day></DateCompleted><DateRevised><Year>2006</Year><Month>11</Month><Day>15</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0167-7012</ISSN><JournalIssue CitedMedium="Print"><Volume>60</Volume><Issue>1</Issue><PubDate><Year>2005</Year><Month>Jan</Month></PubDate></JournalIssue><Title>Journal of microbiological methods</Title><ISOAbbreviation>J. Microbiol. Methods</ISOAbbreviation></Journal><ArticleTitle>Direct detection of Staphylococcus aureus mRNA using a flow through microarray.</ArticleTitle><Pagination><MedlinePgn>47-54</MedlinePgn></Pagination><Abstract><AbstractText>The direct detection of mRNAs from bacterial cultures on a DNA array without amplification and labelling would greatly extend the range of applications suitable for microarray analysis. Here we describe the direct detection of 23S rRNA and seven mRNA species from total Staphylococcus aureus RNA prepared using commercially available RNA purification columns followed by fluorescent detection on a flow through microarray. RNA hybridisation was detected using paired secondary labelled probes directly 5' and 3' to immobilised 60 mers. In this way, we were able to detect the effect of 30-min exposure to antimicrobials on mRNA levels within 3 h after column purification of total RNA without the need for enzymatic manipulation. 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