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Article: Rapid detection of six types of bacterial pathogens in marine waters by multiplex PCR

TitleRapid detection of six types of bacterial pathogens in marine waters by multiplex PCR
Authors
KeywordsBacteria
Marine waters
Multiplex PCR
Pathogen
Virulence
Issue Date2002
PublisherPergamon. The Journal's web site is located at http://www.elsevier.com/locate/watres
Citation
Water Research, 2002, v. 36 n. 11, p. 2802-2812 How to Cite?
AbstractA rapid multiplex PCR (m-PCR) method that allows the simultaneous detection, in a single tube, of six commonly encountered waterborne pathogens is developed. The target genes used were: the aerolysin (aero) gene of Aeromonas hydrophila, the invasion plasmid antigen H (ipaH) gene of Shigella flexneri, the attachment invasion locus (ail) gene of Yersinia enterocolitca, the invasion plasmid antigen B (ipaB) gene of Salmonella typhimurium, the enterotoxin extracellular secretion protein (epsM) gene of Vibrio cholerae and a species-specific region of the 16S-23S rDNA (Vpara) gene of Vibrio parahaemolyticus were used as the gene targets. Multiplex PCR using the six pairs of primers produced specific amplicons of the expected sizes from mixed populations of reference bacterial strains in seawater and from pure cultures. The m-PCR assay was specific and rapid, with a turnaround time of <12h. The detection limit of the assay for the bacterial targets was estimated at 100-102cfu. Multiplex PCR analysis was performed on 19 seawater samples collected around Hong Kong and the results indicated significant levels of four bacterial pathogens at several sites where primary sewage wastes are discharged, and the levels of which showed no correlation with E. coli counts. Overall, both laboratory and field validation results demonstrated that the m-PCR assay developed in this study could provide a cost-effective and informative supplement to conventional microbiological methods for routine monitoring and risk assessment of water quality. © 2002 Elsevier Science Ltd. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/92784
ISSN
2023 Impact Factor: 11.4
2023 SCImago Journal Rankings: 3.596
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorKong, RYCen_HK
dc.contributor.authorLee, SKYen_HK
dc.contributor.authorLaw, TWFen_HK
dc.contributor.authorLaw, SHWen_HK
dc.contributor.authorWu, RSSen_HK
dc.date.accessioned2010-09-17T10:57:03Z-
dc.date.available2010-09-17T10:57:03Z-
dc.date.issued2002en_HK
dc.identifier.citationWater Research, 2002, v. 36 n. 11, p. 2802-2812en_HK
dc.identifier.issn0043-1354en_HK
dc.identifier.urihttp://hdl.handle.net/10722/92784-
dc.description.abstractA rapid multiplex PCR (m-PCR) method that allows the simultaneous detection, in a single tube, of six commonly encountered waterborne pathogens is developed. The target genes used were: the aerolysin (aero) gene of Aeromonas hydrophila, the invasion plasmid antigen H (ipaH) gene of Shigella flexneri, the attachment invasion locus (ail) gene of Yersinia enterocolitca, the invasion plasmid antigen B (ipaB) gene of Salmonella typhimurium, the enterotoxin extracellular secretion protein (epsM) gene of Vibrio cholerae and a species-specific region of the 16S-23S rDNA (Vpara) gene of Vibrio parahaemolyticus were used as the gene targets. Multiplex PCR using the six pairs of primers produced specific amplicons of the expected sizes from mixed populations of reference bacterial strains in seawater and from pure cultures. The m-PCR assay was specific and rapid, with a turnaround time of <12h. The detection limit of the assay for the bacterial targets was estimated at 100-102cfu. Multiplex PCR analysis was performed on 19 seawater samples collected around Hong Kong and the results indicated significant levels of four bacterial pathogens at several sites where primary sewage wastes are discharged, and the levels of which showed no correlation with E. coli counts. Overall, both laboratory and field validation results demonstrated that the m-PCR assay developed in this study could provide a cost-effective and informative supplement to conventional microbiological methods for routine monitoring and risk assessment of water quality. © 2002 Elsevier Science Ltd. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherPergamon. The Journal's web site is located at http://www.elsevier.com/locate/watresen_HK
dc.relation.ispartofWater Researchen_HK
dc.subjectBacteriaen_HK
dc.subjectMarine watersen_HK
dc.subjectMultiplex PCRen_HK
dc.subjectPathogenen_HK
dc.subjectVirulenceen_HK
dc.titleRapid detection of six types of bacterial pathogens in marine waters 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/S0043-1354(01)00503-6en_HK
dc.identifier.pmid12146868-
dc.identifier.scopuseid_2-s2.0-0035987733en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0035987733&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume36en_HK
dc.identifier.issue11en_HK
dc.identifier.spage2802en_HK
dc.identifier.epage2812en_HK
dc.identifier.isiWOS:000176940600015-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridKong, RYC=7005290687en_HK
dc.identifier.scopusauthoridLee, SKY=7601395742en_HK
dc.identifier.scopusauthoridLaw, TWF=7004634665en_HK
dc.identifier.scopusauthoridLaw, SHW=7202242024en_HK
dc.identifier.scopusauthoridWu, RSS=7402945079en_HK
dc.identifier.issnl0043-1354-

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