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Article: Identification of oligonucleotide primers targeted at the 16S-23S rDNA intergenic spacers for genus- and species-specific detection of aeromonads

TitleIdentification of oligonucleotide primers targeted at the 16S-23S rDNA intergenic spacers for genus- and species-specific detection of aeromonads
Authors
Keywords16S-23S rDNA
Aeromonas
Intergenic spacers
PCR
Issue Date1999
PublisherPergamon. The Journal's web site is located at http://www.elsevier.com/locate/marpolbul
Citation
Marine Pollution Bulletin, 1999, v. 38 n. 9, p. 802-808 How to Cite?
AbstractAeromonads occur widely in coastal waters and wastewater; and have been associated with a wide variety of gastrointestinal infections in humans. The prevalence of Aeromonas spp. in the aquatic environment has been recognized as a potential health risk, and some countries have adopted aeromonad counts as an additional indicator of water quality. In view of the above, we have attempted to develop some species-specific primers for the rapid detection of these organisms by PCR. The 16S-23S rRNA intergenic spacer regions (ISR) of seven Aeromonas species - Aeromonas hydrophila, A. caviae, A. enteropelogenes, A. trota, A. jandaei, A. veronii and A. schubertii were amplified by the Polymerase chain reaction using primers complementary to conserved regions of the 16S rRNA and 23S rRNA genes. The ISR amplimers were cloned into plasmid vectors and sequenced. Analysis of the ISR sequences showed that aeromonads contain two types of polymorphic 16S-23S spacers - ISR(Glu) and ISR(IA). The ISR(Glu) contains the tRNA gene for glutamate while ISR(IA) contains the tRNA genes for isoleucine and alanine. Multiple alignment of representative ISR(Glu) sequences from the seven species revealed several major regions of variability. These regions were used to design Aeromonas genus-specific and species-specific primers for PCR. The specificity of the PCR primers were validated using total DNA prepared from various Aeromonas spp. and bacterial species of other genera. Results also showed that the method can be applied efficiently for routine monitoring of aeromonads in seawater Copyright (C) 1999 Elsevier Science Ltd.
Persistent Identifierhttp://hdl.handle.net/10722/92681
ISSN
2023 Impact Factor: 5.3
2023 SCImago Journal Rankings: 1.445
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorKong, RYCen_HK
dc.contributor.authorPelling, Aen_HK
dc.contributor.authorSo, CLen_HK
dc.contributor.authorWu, RSSen_HK
dc.date.accessioned2010-09-17T10:54:01Z-
dc.date.available2010-09-17T10:54:01Z-
dc.date.issued1999en_HK
dc.identifier.citationMarine Pollution Bulletin, 1999, v. 38 n. 9, p. 802-808en_HK
dc.identifier.issn0025-326Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/92681-
dc.description.abstractAeromonads occur widely in coastal waters and wastewater; and have been associated with a wide variety of gastrointestinal infections in humans. The prevalence of Aeromonas spp. in the aquatic environment has been recognized as a potential health risk, and some countries have adopted aeromonad counts as an additional indicator of water quality. In view of the above, we have attempted to develop some species-specific primers for the rapid detection of these organisms by PCR. The 16S-23S rRNA intergenic spacer regions (ISR) of seven Aeromonas species - Aeromonas hydrophila, A. caviae, A. enteropelogenes, A. trota, A. jandaei, A. veronii and A. schubertii were amplified by the Polymerase chain reaction using primers complementary to conserved regions of the 16S rRNA and 23S rRNA genes. The ISR amplimers were cloned into plasmid vectors and sequenced. Analysis of the ISR sequences showed that aeromonads contain two types of polymorphic 16S-23S spacers - ISR(Glu) and ISR(IA). The ISR(Glu) contains the tRNA gene for glutamate while ISR(IA) contains the tRNA genes for isoleucine and alanine. Multiple alignment of representative ISR(Glu) sequences from the seven species revealed several major regions of variability. These regions were used to design Aeromonas genus-specific and species-specific primers for PCR. The specificity of the PCR primers were validated using total DNA prepared from various Aeromonas spp. and bacterial species of other genera. Results also showed that the method can be applied efficiently for routine monitoring of aeromonads in seawater Copyright (C) 1999 Elsevier Science Ltd.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.subject16S-23S rDNAen_HK
dc.subjectAeromonasen_HK
dc.subjectIntergenic spacersen_HK
dc.subjectPCRen_HK
dc.titleIdentification of oligonucleotide primers targeted at the 16S-23S rDNA intergenic spacers for genus- and species-specific detection of aeromonadsen_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/S0025-326X(99)00044-2en_HK
dc.identifier.scopuseid_2-s2.0-0032845918en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0032845918&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume38en_HK
dc.identifier.issue9en_HK
dc.identifier.spage802en_HK
dc.identifier.epage808en_HK
dc.identifier.isiWOS:000083318600018-
dc.publisher.placeUnited Kingdomen_HK
dc.identifier.scopusauthoridKong, RYC=7005290687en_HK
dc.identifier.scopusauthoridPelling, A=6602962712en_HK
dc.identifier.scopusauthoridSo, CL=7102919929en_HK
dc.identifier.scopusauthoridWu, RSS=7402945079en_HK
dc.identifier.issnl0025-326X-

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