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Article: Development of novel oligonucleotide probes for seven Actinomyces species and their utility in supragingival plaque analysis

TitleDevelopment of novel oligonucleotide probes for seven Actinomyces species and their utility in supragingival plaque analysis
Authors
KeywordsActinomyces
Bacterial dierentiation
Hybridization
Oligonucleotide probes
Supragingival plaque
Issue Date2003
PublisherWiley-Blackwell Publishing, Inc.. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=1354-523X&site=1
Citation
Oral Diseases, 2003, v. 9 n. 4, p. 203-209 How to Cite?
AbstractOBJECTIVE: The traditional, biochemical and enzymatic methods of identifying Actinomyces species are frequently confounded by the similar phenotypic characteristics shared by the different members of this genus. Therefore, we developed novel species-specific oligonucleotide probes to accurately speciate seven pathogenic Actinomyces species, namely, Actinomyces bovis, A. gerencseriae, A. israelii, A. meyeri, A. naeslundii, A. odontolyticus and A. viscosus. METHODS: A pair of universal primers and seven 15- to 19-base oligonucleotide probes with a tail of 20 thymidines on the 5′ end were developed. The variable regions of 16S ribosomal DNA of 36 strains of Actinomyces belonging to the above species were amplified and labeled with digoxigenin, and an oligonucleotide-DNA hybridization assay was performed to examine the specificity and sensitivity of these probes. RESULTS: All seven, newly developed probes were specific and sensitive, and accurately detected 36 reference and wild type strains belonging to Actinomyces species, without cross-reactions. The probe for A. naeslundii detected all strains belonging to the genospecies 1 (12 strains) and catalase-negative genospecies 2 (four strains); it failed to detect catalase-positive A. naeslundii genospecies 2 (previous A. viscosus serotype II) (two strains). However, the latter strains of catalase-positive A. naeslundii genospecies 2 were correctly detected by the probe developed for A. viscosus. The new probes were then field tested using supragingival plaque samples from 28 healthy preschool children. Whilst A. odontolyticus was detected in almost all samples (96.4%), A. gerencseriae, A meyeri, catalase-negative A. naeslundii and catalase-positive A. naeslundii genospecies 2 were detected in <50% samples. CONCLUSION: We conclude that the developed oligonucleotide probes, complementary to the variable regions of 16S rDNA, would be of potential value for differentiating Actinomyces spp. in clinical samples from the oral cavity and other ecosystems where such species may abound.
Persistent Identifierhttp://hdl.handle.net/10722/66065
ISSN
2015 Impact Factor: 2.0
2015 SCImago Journal Rankings: 0.828
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorTang, Gen_HK
dc.contributor.authorYip, HKen_HK
dc.contributor.authorLuo, Gen_HK
dc.contributor.authorCheung, BPKen_HK
dc.contributor.authorShen, Sen_HK
dc.contributor.authorSamaranayake, LPen_HK
dc.date.accessioned2010-09-06T05:43:17Z-
dc.date.available2010-09-06T05:43:17Z-
dc.date.issued2003en_HK
dc.identifier.citationOral Diseases, 2003, v. 9 n. 4, p. 203-209en_HK
dc.identifier.issn1354-523Xen_HK
dc.identifier.urihttp://hdl.handle.net/10722/66065-
dc.description.abstractOBJECTIVE: The traditional, biochemical and enzymatic methods of identifying Actinomyces species are frequently confounded by the similar phenotypic characteristics shared by the different members of this genus. Therefore, we developed novel species-specific oligonucleotide probes to accurately speciate seven pathogenic Actinomyces species, namely, Actinomyces bovis, A. gerencseriae, A. israelii, A. meyeri, A. naeslundii, A. odontolyticus and A. viscosus. METHODS: A pair of universal primers and seven 15- to 19-base oligonucleotide probes with a tail of 20 thymidines on the 5′ end were developed. The variable regions of 16S ribosomal DNA of 36 strains of Actinomyces belonging to the above species were amplified and labeled with digoxigenin, and an oligonucleotide-DNA hybridization assay was performed to examine the specificity and sensitivity of these probes. RESULTS: All seven, newly developed probes were specific and sensitive, and accurately detected 36 reference and wild type strains belonging to Actinomyces species, without cross-reactions. The probe for A. naeslundii detected all strains belonging to the genospecies 1 (12 strains) and catalase-negative genospecies 2 (four strains); it failed to detect catalase-positive A. naeslundii genospecies 2 (previous A. viscosus serotype II) (two strains). However, the latter strains of catalase-positive A. naeslundii genospecies 2 were correctly detected by the probe developed for A. viscosus. The new probes were then field tested using supragingival plaque samples from 28 healthy preschool children. Whilst A. odontolyticus was detected in almost all samples (96.4%), A. gerencseriae, A meyeri, catalase-negative A. naeslundii and catalase-positive A. naeslundii genospecies 2 were detected in <50% samples. CONCLUSION: We conclude that the developed oligonucleotide probes, complementary to the variable regions of 16S rDNA, would be of potential value for differentiating Actinomyces spp. in clinical samples from the oral cavity and other ecosystems where such species may abound.en_HK
dc.languageengen_HK
dc.publisherWiley-Blackwell Publishing, Inc.. The Journal's web site is located at http://www.wiley.com/bw/journal.asp?ref=1354-523X&site=1en_HK
dc.relation.ispartofOral Diseasesen_HK
dc.subjectActinomycesen_HK
dc.subjectBacterial dierentiationen_HK
dc.subjectHybridizationen_HK
dc.subjectOligonucleotide probesen_HK
dc.subjectSupragingival plaqueen_HK
dc.subject.mesh5' Flanking Regionen_HK
dc.subject.meshActinomyces - classification - geneticsen_HK
dc.subject.meshActinomyces viscosus - classification - geneticsen_HK
dc.subject.meshChild, Preschoolen_HK
dc.subject.meshDNA, Bacterial - analysis - geneticsen_HK
dc.subject.meshDental Plaque - microbiologyen_HK
dc.subject.meshHumansen_HK
dc.subject.meshNucleic Acid Hybridizationen_HK
dc.subject.meshOligonucleotide Probes - diagnostic useen_HK
dc.subject.meshRNA, Ribosomal, 16S - geneticsen_HK
dc.subject.meshSensitivity and Specificityen_HK
dc.subject.meshSerotypingen_HK
dc.subject.meshSpecies Specificityen_HK
dc.subject.meshThymidine - diagnostic useen_HK
dc.titleDevelopment of novel oligonucleotide probes for seven Actinomyces species and their utility in supragingival plaque analysisen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1354-523X&volume=9&spage=203&epage=209&date=2003&atitle=Development+of+Novel+Oligonucleotide+Probes+for+Seven+Actinomyces+Species+and+Their+Utility+in+Supragingival+Plaque+Analysisen_HK
dc.identifier.emailYip, HK: kevin.h.k.yip@hkusua.hku.hken_HK
dc.identifier.emailSamaranayake, LP: lakshman@hku.hken_HK
dc.identifier.authorityYip, HK=rp00027en_HK
dc.identifier.authoritySamaranayake, LP=rp00023en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1034/j.1601-0825.2003.02926.xen_HK
dc.identifier.pmid12974520-
dc.identifier.scopuseid_2-s2.0-0038721780en_HK
dc.identifier.hkuros77255en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0038721780&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume9en_HK
dc.identifier.issue4en_HK
dc.identifier.spage203en_HK
dc.identifier.epage209en_HK
dc.identifier.isiWOS:000183515300006-
dc.publisher.placeUnited Statesen_HK
dc.identifier.scopusauthoridTang, G=36882453100en_HK
dc.identifier.scopusauthoridYip, HK=25423244900en_HK
dc.identifier.scopusauthoridLuo, G=55112399700en_HK
dc.identifier.scopusauthoridCheung, BPK=7103294773en_HK
dc.identifier.scopusauthoridShen, S=7403431743en_HK
dc.identifier.scopusauthoridSamaranayake, LP=7102761002en_HK
dc.identifier.citeulike3010911-

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