[Rapid detection of the Escherichia cori verotoxin gene using fluorescence polarization].
Identifieur interne : 003C02 ( Main/Exploration ); précédent : 003C01; suivant : 003C03[Rapid detection of the Escherichia cori verotoxin gene using fluorescence polarization].
Auteurs : M. Tsuruoka ; T. Honda ; I. KarubeSource :
- Nihon rinsho. Japanese journal of clinical medicine [ 0047-1852 ] ; 1997.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : DNA, Bacterial.
- chemical , genetics : Bacterial Toxins.
- genetics : Escherichia coli O157.
- Fluorescence Polarization, Nucleic Acid Hybridization, Shiga Toxin 1, Time Factors.
Abstract
The effects of NaCl concentration, temperature and base-pair mismatches on the hybridization of two complementary single-stranded DNA 24-mers were investigated using a fluorescence polarization method. Over a temperature range of 46 degrees C to 56 degrees C in 0.8 M NaCl it was found that hybridization was essentially complete in under 10 minutes and that rapid in vitro determination of the target DNA was possible when the sample DNA had three or less base-pair mismatches in the 24-mer sequence. Polarization measurements on positive and negative samples showed excellent agreement with results obtained from electrophoresis. Under our optimized conditions, a 23 base-pair single-stranded DNA sequence of the Verotoxin gene (VT2) of Escherichia coli, previously multiplied using PCR (40 cycles), could be detected within 10 minutes.
PubMed: 9086791
Affiliations:
Links toward previous steps (curation, corpus...)
- to stream PubMed, to step Corpus: 002734
- to stream PubMed, to step Curation: 002734
- to stream PubMed, to step Checkpoint: 002561
- to stream Ncbi, to step Merge: 002A69
- to stream Ncbi, to step Curation: 002A69
- to stream Ncbi, to step Checkpoint: 002A69
- to stream Main, to step Merge: 003C53
- to stream Main, to step Curation: 003C02
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">[Rapid detection of the Escherichia cori verotoxin gene using fluorescence polarization].</title>
<author><name sortKey="Tsuruoka, M" sort="Tsuruoka, M" uniqKey="Tsuruoka M" first="M" last="Tsuruoka">M. Tsuruoka</name>
<affiliation><nlm:affiliation>Advanced Science and Technology Laboratory, City of Hiroshima.</nlm:affiliation>
<wicri:noCountry code="subField">City of Hiroshima</wicri:noCountry>
</affiliation>
</author>
<author><name sortKey="Honda, T" sort="Honda, T" uniqKey="Honda T" first="T" last="Honda">T. Honda</name>
</author>
<author><name sortKey="Karube, I" sort="Karube, I" uniqKey="Karube I" first="I" last="Karube">I. Karube</name>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="1997">1997</date>
<idno type="RBID">pubmed:9086791</idno>
<idno type="pmid">9086791</idno>
<idno type="wicri:Area/PubMed/Corpus">002734</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">002734</idno>
<idno type="wicri:Area/PubMed/Curation">002734</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">002734</idno>
<idno type="wicri:Area/PubMed/Checkpoint">002561</idno>
<idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">002561</idno>
<idno type="wicri:Area/Ncbi/Merge">002A69</idno>
<idno type="wicri:Area/Ncbi/Curation">002A69</idno>
<idno type="wicri:Area/Ncbi/Checkpoint">002A69</idno>
<idno type="wicri:doubleKey">0047-1852:1997:Tsuruoka M:rapid:detection:of</idno>
<idno type="wicri:Area/Main/Merge">003C53</idno>
<idno type="wicri:Area/Main/Curation">003C02</idno>
<idno type="wicri:Area/Main/Exploration">003C02</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">[Rapid detection of the Escherichia cori verotoxin gene using fluorescence polarization].</title>
<author><name sortKey="Tsuruoka, M" sort="Tsuruoka, M" uniqKey="Tsuruoka M" first="M" last="Tsuruoka">M. Tsuruoka</name>
<affiliation><nlm:affiliation>Advanced Science and Technology Laboratory, City of Hiroshima.</nlm:affiliation>
<wicri:noCountry code="subField">City of Hiroshima</wicri:noCountry>
</affiliation>
</author>
<author><name sortKey="Honda, T" sort="Honda, T" uniqKey="Honda T" first="T" last="Honda">T. Honda</name>
</author>
<author><name sortKey="Karube, I" sort="Karube, I" uniqKey="Karube I" first="I" last="Karube">I. Karube</name>
</author>
</analytic>
<series><title level="j">Nihon rinsho. Japanese journal of clinical medicine</title>
<idno type="ISSN">0047-1852</idno>
<imprint><date when="1997" type="published">1997</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Bacterial Toxins (genetics)</term>
<term>DNA, Bacterial (analysis)</term>
<term>Escherichia coli O157 (genetics)</term>
<term>Fluorescence Polarization</term>
<term>Nucleic Acid Hybridization</term>
<term>Shiga Toxin 1</term>
<term>Time Factors</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>ADN bactérien (analyse)</term>
<term>Escherichia coli O157 (génétique)</term>
<term>Facteurs temps</term>
<term>Hybridation d'acides nucléiques</term>
<term>Polarisation de fluorescence</term>
<term>Shiga-toxine-1</term>
<term>Toxines bactériennes (génétique)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>DNA, Bacterial</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Bacterial Toxins</term>
</keywords>
<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>ADN bactérien</term>
</keywords>
<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Escherichia coli O157</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Escherichia coli O157</term>
<term>Toxines bactériennes</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Fluorescence Polarization</term>
<term>Nucleic Acid Hybridization</term>
<term>Shiga Toxin 1</term>
<term>Time Factors</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Facteurs temps</term>
<term>Hybridation d'acides nucléiques</term>
<term>Polarisation de fluorescence</term>
<term>Shiga-toxine-1</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">The effects of NaCl concentration, temperature and base-pair mismatches on the hybridization of two complementary single-stranded DNA 24-mers were investigated using a fluorescence polarization method. Over a temperature range of 46 degrees C to 56 degrees C in 0.8 M NaCl it was found that hybridization was essentially complete in under 10 minutes and that rapid in vitro determination of the target DNA was possible when the sample DNA had three or less base-pair mismatches in the 24-mer sequence. Polarization measurements on positive and negative samples showed excellent agreement with results obtained from electrophoresis. Under our optimized conditions, a 23 base-pair single-stranded DNA sequence of the Verotoxin gene (VT2) of Escherichia coli, previously multiplied using PCR (40 cycles), could be detected within 10 minutes.</div>
</front>
</TEI>
<affiliations><list></list>
<tree><noCountry><name sortKey="Honda, T" sort="Honda, T" uniqKey="Honda T" first="T" last="Honda">T. Honda</name>
<name sortKey="Karube, I" sort="Karube, I" uniqKey="Karube I" first="I" last="Karube">I. Karube</name>
<name sortKey="Tsuruoka, M" sort="Tsuruoka, M" uniqKey="Tsuruoka M" first="M" last="Tsuruoka">M. Tsuruoka</name>
</noCountry>
</tree>
</affiliations>
</record>
Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/MersV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 003C02 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 003C02 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien |wiki= Sante |area= MersV1 |flux= Main |étape= Exploration |type= RBID |clé= pubmed:9086791 |texte= [Rapid detection of the Escherichia cori verotoxin gene using fluorescence polarization]. }}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:9086791" \ | HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \ | NlmPubMed2Wicri -a MersV1
![]() | This area was generated with Dilib version V0.6.33. | ![]() |