Detection of Actinobacillus, actinomycetemcomitans by culture methods and multiplex PCR in sub gingival plaque samples of Sudanese aggressive periodontitis patients

by Open Science Repository Dentistry
(June 2014)

Abstract

Aggressive periodontitis is a destructive priodontal disease which may lead to early loss of teeth. The etiology of the disease is unknown but there are many studies about the implication of gram-negative bacteria Actinobacillus actinomycetumcomitans, Prophronomonasgingivalis, P.intermedia and T.Fosyrthia the disease progression, also there is a genetic factor ,(defect in polymorph nuclear leucocytes) that may be involved. This study is done to identify the major causative micro-organism Actinobacillus actinomycetumcomitans, by culture and multiplex PCR.

Method: Thirty aggressive periodontitis patients were recruited from patients attending department of periodontology ,Faculty of dentistry, University of Khartoum for treatment. The age range 18-35 years, both male and female. Sub-gingival plaque samples were obtained from two deepest site per quadrant for each patient for identification of the major causative micro-organisms Actinobacillus actinomycetumcomitans. Identification was done by anaerobic culture in blood agar and sub-cultured using vancomycin. After DNA extraction multiplex PCR technique which is more sensitive was used for confirmation and to identify serotypes of Actinobacillus actinomycetumcomitans if present in the examined samples.

Results: Actinobacillus actinomycetumcomitans was not identified in all examined sites by culture and this was confirmed with biochemical testes.Using  multiplex PCR and after electrophoreses  no bands were amplified from all samples .The two methods showed identical results in the identification of Actinobacillus actinomycetumcomitans.

Conclusion: Since A. actinomycetemcomitan was not present in the examined samples using both culture and multiplex PCR ,so its role in the pathogeneses of the disease may be questionable.

Keywords: Actinobacillus actinomycetumcomitans, Multiplex PCR, culture.


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Detection of Actinobacillus, actinomycetemcomitans by culture methods and multiplex PCR in sub gingival plaque samples of Sudanese aggressive periodontitis patients

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