Identification of Veillonella spp. on Tongue Plaque and Saliva Using Real-Time PCR

https://doi.org/10.22146/theindjdentres.65715

Karina Dhaniarti(1), Fathia Agzarine Deandra(2*), Ariadna A Djais(3), Boy M Bachtiar(4)

(1) Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta
(2) Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta
(3) Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta
(4) Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta
(*) Corresponding Author

Abstract


Veillonella spp., Gram-negative obligate anaerobic cocci bacteria, amounts to 3% in the oral cavity, relies on the fermentation of lactate as a carbon and energy source for growth. The bacteria are considered anti-cariogenic as they metabolize lactic acid into propionic acid which increases oral environment’s pH and reduces demineralization rate of tooth structure. Identification of Veillonella spp. using traditional methods is difficult due to the lack of conventional phenotypic and biochemical tests. Thus, the biomolecular methods are suitable for the specific detection and identification of Veillonella spp. One of the biomolecular methods that can be used is real-time Polymerase Chain Reaction (PCR), which the results can be qualitative and quantitative. This study aimed to identify Veillonella spp. in tongue plaque’s and saliva’s samples using Real -time PCR. The DNA of Veillonella spp. derived from 36 samples, 18 samples of tongue plaque and 18 samples of saliva, were extracted using a freeze-thaw method and then quantified by real-time PCR using forward primer 5’-CCG TGA TGG GAT GGA AAC TGC-3’ and reverse primer 5’-CCT TCG CCA CTG GTG TTC TTC-3’. Veillonella spp. in 18 samples of tongue plaque was 3,06 x 107 CFU/ml and in 18 saliva samples was 1,51 x 105 CFU/ml.  It was concluded real-time PCR can detect Veillonella spp. from all tongue plaque’s and saliva’s samples.
 


Keywords


Veillonella spp.; real-time PCR; tongue plaque; saliva

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References

Samaranayake L. Essential Microbiology for Dentistry. 4th ed.

Edinburgh: Churchill Livingstone; 2012. 140 p.

Mashima I, Futoshi Nakazawa. Identification of Veillonella

tobetsuensis in Tongue Biofilm by Using A Species-Specific Primer Pair. Elsevier. 2013;22:77–81.

Mashima I, Futoshi Nakazawa. A Review on the Characterization of A Novel Oral Veillonella Species, V. tobetsuensis, and Its Role in Biofilm Formation. Elsevier. 2013;55:184–90.

Byun R, Jean-Philippe Carlier, Nicholas A. Jacques, Helene Marchandin, Neil Hunter. Veillonella denticariosi sp.nov., Isolated from Human Carious Dentine. IJSEM. 2007;57(12):2844–8.

Joshi DM, Dr. Deshpande J.D. Polymerase Chain Reaction: Methods, Principles and Application. Int J Biomed Res. 2010;1(5):81–97.

Life Technologies. Real-Time PCR Handbook [Internet]. [cited 2014 Oct 12]. Available from: http://find.lifetechnologies.com/Global/FileLib/qPCR/RealTimePCR_Handbook_Update_FLR.pdf

Bio-Rad. Real-Time PCR: Applications Guide [Internet]. [cited 2014 Oct 12]. Available from: http://www.biorad.com/webroot/web/pdf/lsr/literature/Bulletin_5279.pdf

AAPD, Council on Clinical Affairs. AAPD Caries risk assessment tool (CAT)* [Internet]. 2014 [cited 2014 Oct 8].

Available from: http://www.uhccommunityplan.com/content/dam/communityplan/healthcareprofessionals/providerinformation

/RiskAssesmentTool.pdf

Mashima I, Kamaguchi A, Nakazawa F. The Distribution and Frequency of Oral Veillonella spp. in Tongue Biofim of Healthy Young Adults. Curr Microbiol. 2011;63(5):403–7.

PCR [Internet]. Probe Reagents for Functional Genomics. [cited 2014 Oct 13]. Available from: http://www.ncbi.nlm.nih.gov/projects/genome/probe/doc/TechPCR.shtml

Principle of the PCR [Internet]. [cited 2014 Oct 13]. Available from: http://users.ugent.be/~avierstr/principles/pcr.html

Arif N, E. C. Sheehy, T. Do, D Beighton. Diversity of Veillonella spp. from Sound and Carious Sites in Children. PMC. 2008 Mar 10;87(3):278–82.

Premier Biosoft. Identifying and distinguishing bacterial strains using Real Time PCR and Microarrays [Internet]. Identifying Bacteria. Available from: http://www.premierbiosoft.com/tech

_notes/bac-id.html

Total DNA Extraction by Freeze-Thaw [Internet]. [cited 2015 Jan 19]. Available from: http://webs.wichita.edu/mschneegurt

/gk12web/classroom/protocols/freezethaw.doc

Andree KB, Margarita FernandezTejedor, Laurence M. Elandaloussi, et al. Quantitative PCR Coupled with Melt Curve Analysis for Detectioin of Selected Pseudonitzschia spp. (Bacillariophyceae) from the Northwestern Mediterranean Sea.

Mar;77(5):1651–9.

Matsui M, Naoyuki Chosa, Yu Shimoyama, et al. Effects of Tongue Cleaning on Bacterial Flora in Tongue Coating and Dental Plaque: A Crossover Study. BMC Oral Health [Internet]. 2014;14(4). Available from: http://www.biomedcentral.com/content/pdf/1472-6831-14-4.pdf

Kreth J, Justin Merritt, Fengxia Qi. Bacterial and Host Interactions of OralStreptococci. PMC. 2009Aug;28(8):397–403.

Winnier JJ, Ullal Anand Nayak,Venugopal Reddy, Arun Prasad Rao. The Comparative Evaluation of theEffects of Tongue Cleaning on ExistingPlaque Levels in Chilren. PMC.2013;6(3):188–92.



DOI: https://doi.org/10.22146/theindjdentres.65715

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