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

Tang, G., Yip, H. -K., Luo, G., Cheung, B. P. K., Shen, S. and Samaranayake, L. P. (2003) Development of novel oligonucleotide probes for seven Actinomyces species and their utility in supragingival plaque analysis. Oral Diseases, 9 4: 203-209. doi:10.1034/j.1601-0825.2003.02926.x


Author Tang, G.
Yip, H. -K.
Luo, G.
Cheung, B. P. K.
Shen, S.
Samaranayake, L. P.
Title Development of novel oligonucleotide probes for seven Actinomyces species and their utility in supragingival plaque analysis
Formatted title
Development of novel oligonucleotide probes for seven Actinomyces species and their utility in supragingival plaque analysis
Journal name Oral Diseases   Check publisher's open access policy
ISSN 1354-523X
1601-0825
Publication date 2003-07-01
Year available 2003
Sub-type Article (original research)
DOI 10.1034/j.1601-0825.2003.02926.x
Open Access Status Not yet assessed
Volume 9
Issue 4
Start page 203
End page 209
Total pages 7
Place of publication Hoboken, NJ, United States
Publisher Wiley-Blackwell Publishing
Language eng
Formatted abstract
Objective: 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.
Keyword Actinomyces
Bacterial dierentiation
Hybridization
Oligonucleotide probes
Supragingival plaque
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: School of Dentistry Publications
 
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