Single multiplex polymerase chain reaction for environmental surveillance of toxigenic-pathogenic O1 and non-O1 Vibrio cholerae
Jazyk angličtina Země Spojené státy americké Médium print
Typ dokumentu hodnotící studie, časopisecké články
PubMed
17571801
DOI
10.1007/bf02932143
Knihovny.cz E-zdroje
- MeSH
- bakteriální proteiny genetika MeSH
- cholerový toxin genetika MeSH
- DNA primery MeSH
- látky znečišťující vodu analýza MeSH
- monitorování životního prostředí metody MeSH
- polymerázová řetězová reakce metody MeSH
- senzitivita a specificita MeSH
- sladká voda mikrobiologie MeSH
- Vibrio cholerae non-O1 * genetika izolace a purifikace patogenita MeSH
- Vibrio cholerae O1 * genetika izolace a purifikace patogenita MeSH
- zásobování vodou analýza MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- Názvy látek
- bakteriální proteiny MeSH
- cholerový toxin MeSH
- DNA primery MeSH
- látky znečišťující vodu MeSH
A multiplex PCR assay was developed for the detection of toxigenic and pathogenic V. cholerae from direct water sources using specific primers targeting diverse genes, viz. outer membrane protein (ompW), cholera toxin (ctxB), ORF specific for O1 (rfbG), zonula occludens (zot) and toxin co-regulated pilus (tcpB); among these genes, ompW acts as internal control for V. cholerae, the ctx gene as a marker for toxigenicity and tcp for pathogenicity. The sensitivity of multiplex PCR was 5 x 10(4) V. cholerae cells per reaction. The procedure was simplified as direct bacterial cells were used as template and there was no need for DNA extraction. The assay was specific as no amplification occurred with the other bacteria used. Toxigenic V. cholerae were artificially spiked in different water samples, filtered through a 0.45 microm membrane, and the filters containing bacteria were enriched in APW for 6 h. PCR following filtration and enrichment could detect as little as 8 V. cholerae cells per mL in different spiked water samples. Various environmental potable water samples were screened for the presence of V. cholerae using this assay procedure. The proposed method is rapid, sensitive and specific for environmental surveillance for the presence of toxigenic-pathogenic and nonpathogenic V. cholerae.
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