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Article: Co-detection of three species of water-borne bacteria by multiplex PCR

TitleCo-detection of three species of water-borne bacteria by multiplex PCR
Authors
KeywordsBacterium Contamination
Controlled Study
Diagnostic Accuracy
Dna Template
Escherichia Coli
Housekeeping Gene
Nonhuman
Polymerase Chain Reaction
Public Health
Salmonella Typhimurium
Species Difference
Vibrio Cholerae
Water Contamination
Water Quality
Issue Date1995
PublisherPergamon. The Journal's web site is located at http://www.elsevier.com/locate/marpolbul
Citation
Marine Pollution Bulletin, 1995, v. 31 n. 4-12, p. 317-324 How to Cite?
AbstractMonitoring of water-borne pathogens is important to safeguard public health. In view of various limitations inherent in the traditional culture methods, the feasibility of using the polymerase chain reaction (PCR) to monitor water-borne pathogens was investigated. The STN enterotoxin gene of Salmonella typhimurium, the STO enterotoxin gene of Vibrio cholerae, the LTI and LTII enterotoxin genes of Escheuichia coli, and the housekeeping genes, ARO-A and PHO-A of S. typhimurium and E. coli, respectively, were used as gene targets for PCR detection of toxigenic and general strains of these organisms. Six pairs of oligonucleotide primers were chosen to amplify internal fragments of the respective genes, and the identity of the PCR products was confirmed by restriction endonuclease digestion. The specificity of individual primer pairs in PCR was evaluated on DNA templates of 54 different bacterial isolates. The results showed that the LTI, LTII and STO primer sets were highly specific for toxigenic strains of E. coli H10407 (LTI+), E. coli SA53 (LTII+) and V. cholerae NRT (STO+), respectively. The PHO-A primers showed species-specific amplification products for all nine E. coli isolates examined, while the STN and ARO-A primer sets yielded species-specific amplification products for the 10 S. typhimurium isolates tested. Detection sensitivity of the ARO-A and PHO-A primer sets for S. typhlmurium and E. coli, respectively, was estimated at 103 CFU. Using three different combinations of the above primer sets, multiplex PCR was performed to detect toxigenic and non-toxigenic strains of V. cholerae, S. typhimurium and E. coli in seawater samples artificially spiked with the organisms. The technique, which showed positive co-detection of the respective target genes in each case, only required a turnaround time of 5 h. Results of the present study indicate that the multiplex PCR is a potentially powerful technique for the rapid codetection of enteropathogenic bacteria in routine water quality monitoring.
Persistent Identifierhttp://hdl.handle.net/10722/92764
ISSN
2015 Impact Factor: 3.099
2015 SCImago Journal Rankings: 1.264
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorKong, RYCen_HK
dc.contributor.authorDung, WFen_HK
dc.contributor.authorVrijmoed, LLPen_HK
dc.contributor.authorWu, RSSen_HK
dc.date.accessioned2010-09-17T10:56:28Z-
dc.date.available2010-09-17T10:56:28Z-
dc.date.issued1995en_HK
dc.identifier.citationMarine Pollution Bulletin, 1995, v. 31 n. 4-12, p. 317-324en_HK
dc.identifier.issn0025-326Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/92764-
dc.description.abstractMonitoring of water-borne pathogens is important to safeguard public health. In view of various limitations inherent in the traditional culture methods, the feasibility of using the polymerase chain reaction (PCR) to monitor water-borne pathogens was investigated. The STN enterotoxin gene of Salmonella typhimurium, the STO enterotoxin gene of Vibrio cholerae, the LTI and LTII enterotoxin genes of Escheuichia coli, and the housekeeping genes, ARO-A and PHO-A of S. typhimurium and E. coli, respectively, were used as gene targets for PCR detection of toxigenic and general strains of these organisms. Six pairs of oligonucleotide primers were chosen to amplify internal fragments of the respective genes, and the identity of the PCR products was confirmed by restriction endonuclease digestion. The specificity of individual primer pairs in PCR was evaluated on DNA templates of 54 different bacterial isolates. The results showed that the LTI, LTII and STO primer sets were highly specific for toxigenic strains of E. coli H10407 (LTI+), E. coli SA53 (LTII+) and V. cholerae NRT (STO+), respectively. The PHO-A primers showed species-specific amplification products for all nine E. coli isolates examined, while the STN and ARO-A primer sets yielded species-specific amplification products for the 10 S. typhimurium isolates tested. Detection sensitivity of the ARO-A and PHO-A primer sets for S. typhlmurium and E. coli, respectively, was estimated at 103 CFU. Using three different combinations of the above primer sets, multiplex PCR was performed to detect toxigenic and non-toxigenic strains of V. cholerae, S. typhimurium and E. coli in seawater samples artificially spiked with the organisms. The technique, which showed positive co-detection of the respective target genes in each case, only required a turnaround time of 5 h. Results of the present study indicate that the multiplex PCR is a potentially powerful technique for the rapid codetection of enteropathogenic bacteria in routine water quality monitoring.en_HK
dc.languageengen_HK
dc.publisherPergamon. The Journal's web site is located at http://www.elsevier.com/locate/marpolbulen_HK
dc.relation.ispartofMarine Pollution Bulletinen_HK
dc.subjectBacterium Contaminationen_HK
dc.subjectControlled Studyen_HK
dc.subjectDiagnostic Accuracyen_HK
dc.subjectDna Templateen_HK
dc.subjectEscherichia Colien_HK
dc.subjectHousekeeping Geneen_HK
dc.subjectNonhumanen_HK
dc.subjectPolymerase Chain Reactionen_HK
dc.subjectPublic Healthen_HK
dc.subjectSalmonella Typhimuriumen_HK
dc.subjectSpecies Differenceen_HK
dc.subjectVibrio Choleraeen_HK
dc.subjectWater Contaminationen_HK
dc.subjectWater Qualityen_HK
dc.titleCo-detection of three species of water-borne bacteria by multiplex PCRen_HK
dc.typeArticleen_HK
dc.identifier.emailWu, RSS: rudolfwu@hku.hken_HK
dc.identifier.authorityWu, RSS=rp01398en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/0025-326X(95)00139-Een_HK
dc.identifier.scopuseid_2-s2.0-0029416220en_HK
dc.identifier.volume31en_HK
dc.identifier.issue4-12en_HK
dc.identifier.spage317en_HK
dc.identifier.epage324en_HK
dc.identifier.isiWOS:A1995TM84200024-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridKong, RYC=7005290687en_HK
dc.identifier.scopusauthoridDung, WF=23066770000en_HK
dc.identifier.scopusauthoridVrijmoed, LLP=7003337180en_HK
dc.identifier.scopusauthoridWu, RSS=7402945079en_HK

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