Abstract Aim The objective of this study was to evaluate the use of unstimulated saliva in detecting Aggregatibacter actinomycetemcomitans and its JP2 genotype, and to compare the saliva, subgingival and mucosa membrane occurrence of this periopathogen in patients diagnosed with advanced periodontitis. Materials and Methods One hundred and fifty-six patients (mean age: 54.03 ± 03 years) with advanced forms of periodontitis (stage-III/IV) were sampled. Unstimulated saliva, buccal cheek mucosa and pooled subgingival plaque samples were collected. Identification of A. actinomycetemcomitans and its JP2 genotype was performed using polymerase chain reaction. A descriptive analysis and chi-square test evaluation were performed. Results A. actinomycetemcomitans was isolated from 32% of the subjects. A total of 468 samples were obtained, 156 from unstimulated saliva, buccal cheek mucosa surfaces, and pooled subgingival plaque samples. A. actinomycetemcomitans was isolated from 23.07% of unstimulated saliva samples, 19.87% of buccal cheek swab samples, and 17.30% of subgingival samples. The total 468 samples were negative for the JP2 genotype of A. actinomycetemcomitans in this cohort of patients. Conclusions These results suggest that in advanced periodontitis, unstimulated saliva is representative of pooled subgingival plaque/buccal cheek samples and its use is adequate in the oral detection of A. actinomycetemcomitans in a cohort of patients with stage-III and IV periodontitis. Clinical Relevance: Data related to the oral colonization of periopthogens can be crucial to the diagnosis, vaccination, maintenance, and treatment of advanced forms of periodontitis. A. actinomycetemcomitans is a key periopathogen implicated in the pathogenesis of periodontitis. In the presence of inflammation, advanced bone loss and deep periodontal pocketing, saliva samples seem to be an effective and conservative alternative to other methods of sampling in detecting the presence of A. actinomycetemcomitans.
The aims of the present study were to document the presence of Aggregatibacter actinomyctemcomitans and the emerging oral pathogen Filifactor alocis, as well as to identify genotypes of these bacterial species with enhanced virulence. In addition, these data were analyzed in relation to periodontal pocket depth (PPD) and the progression of PPD from the sampled periodontal sites during a two-year period. Subgingival plaque samples were collected from 172 periodontal pockets of 68 Ghanaian adolescents. PPD at sampling varied from 3-14 mm and the progression from baseline, i.e., two years earlier up to 8 mm. The levels of A. actinomycetemcomitans and F. alocis were determined with quantitative PCR. The highly leukotoxic JP2-genotype of A. actinomycetemcomitans and the ftxA a gene of F. alocis, encoding a putative Repeats-in-Toxin (RTX) protein, were detected with conventional PCR. The prevalence of A. actinomycetemcomitans was 57%, and 14% of the samples contained the JP2 genotype. F. alocis was detected in 92% of the samples and the ftxA gene in 52%. The levels of these bacterial species were significantly associated with enhanced PPD and progression, with a more pronounced impact in sites positive for the JP2 genotype or the ftxA gene. Taken together, the results indicate that the presence of both A. actinomycetemcomitans and F. alocis with their RTX proteins are linked to increased PPD and progression of disease.
This paper aims to investigate the presence of Aggregatibacter actinomycetemcomitans and to assess potential indicators of the risk of severe form(s) of periodontitis. A descriptive cross-sectional study of 156 consecutive patients with periodontitis was conducted. Subgingival plaque samples were collected from the participants. The identification of A. actinomycetemcomitans was performed using quantitative polymerase chain reaction. A descriptive analysis, a chi-square test, and a binary logistic regression statistical evaluation were performed. The prevalence of A. actinomycetemcomitans in this population of 156 participants was 17.30% (27 patients). The prevalence of stage-III periodontitis was 75.6% and greater in older men, while the prevalence of stage-IV periodontitis was 22.4% and greater in younger women. We observed a significant relation between the risk of severe periodontitis (stage-IV) and poor oral hygiene (p = 0.006), attendance at dental appointments (p ≤ 0.001), and familial history of periodontitis (p = 0.032). In conclusion, twenty-seven individuals were positive for A. actinomycetemcomitans. Poor oral hygiene, family history of periodontitis, and irregular attendance at dental appointments were identified as potential risk factors for severe periodontitis in this cohort.
The Gram-positive bacterium, Filifactor alocis is an oral pathogen, and approximately 50% of known strains encode a recently identified repeat-in-toxin (RTX) protein, FtxA. By assessing a longitudinal Ghanaian study population of adolescents (10-19 years of age; mean age 13.2 years), we recently discovered a possible correlation between deep periodontal pockets measured at the two-year follow-up, presence of the ftxA gene, and a high quantity of F. alocis . To further understand the contribution of F. alocis and FtxA in periodontal disease, we used qPCR in the present study to assess the carriage loads of F. alocis and the prevalence of its ftxA gene in subgingival plaque specimens, sampled at baseline from the Ghanaian cohort (n=500). Comparing these results with the recorded clinical attachment loss (CAL) longitudinal progression data from the two-year follow up, we concluded that carriers of ftxA -positive F. alocis typically exhibited higher loads of the bacterium. Moreover, high carriage loads of F. alocis and concomitant presence of the ftxA gene were two factors that were both associated with an enhanced prevalence of CAL progression. Interestingly, CAL progression appeared to be further promoted upon the simultaneous presence of F. alocis and the non-JP2 genotype of Aggregatibacter actinomycetemcomitans . Taken together, our present findings are consistent with the notion that F. alocis and its ftxA gene promotes CAL during periodontal disease.
The Gram-positive organism Filifactor alocis is implicated in multiple oral diseases including periodontitis, and approximately 50% of known strains encode and produce a recently identified repeat-in-toxin (RTX) protein, FtxA, partly homologous to the Aggregatibacter actinomycetemcomitans leukotoxin. By assessing a longitudinal Ghanaian study population of adolescents, we recently identified a possible correlation between F. alocis levels, ftxA gene carriage, and progression of clinical attachment loss (CAL). To extend knowledge on the possible significance of F. alocis and its FtxA in periodontal disease, we have in the present work analyzed saliva samples in an independent cohort of periodontitis (n=156), collected at two private periodontal specialist practices in Perth, Western Australia. The present results corroborate that high loads of F. alocis and the presence of its ftxA gene together are associated with parameters of periodontal tissue destruction and severity. Moreover, among the individuals carrying A. actinomycetemcomitans , a majority also exhibited an ftxA -positive F. alocis , supporting the notion of the synergistic behavior of these two species. This emphasizes that F. alocis and its ftxA are involved in the pathogenesis of periodontitis and may have ecological roles, with diagnostic and prognostic implications for the disease.
Abstract Aim To investigate the presence of Aggregatibacter actinomycetemcomitans in subgingival plaque samples isolated from patients with periodontitis, and to assess potential indicators of the risk of severe form(s) of periodontitis. Materials and Methods A descriptive cross-sectional study of 156 consecutive patients with periodontitis, aged between 26 and 86 years old, were recruited to two private periodontal practices in Western Australia, between June and November 2022. After collection of the patient’s biodata, periodontal, radiographic and photogenic examination, subgingival plaque samples were collected from shallow and deep periodontal pockets of the participants. Identification of A. actinomycetemcomitans was performed using quantitative Polymerase Chain Reaction utilising species-specific primers. A descriptive analysis, chi-square test, and binary logistic regression statistical evaluation were performed. Results The prevalence of A. actinomycetemcomitans in this population of 156 participants was 16% (25 patients). Using the 2018 Classification of Periodontal and Peri-implant Diseases and Conditions, the prevalence of stage-III periodontitis was 75.6% and more prevalent in older men, while the prevalence of stage-IV periodontitis was 22.4% and more prevalent in younger women. The most prevalent extension of periodontitis was the generalized form with 83.3%. The most common periodontitis grade was B (73.1%). We observed a significant relation of the risk of severe periodontitis (stage-IV) with poor oral hygiene (X 2 = 10.15, p = 0.006), and attendance to dental appointments (X 2 = 14.78, p = < 0.001 - B = 1.40, OR = 4.079; 95% CI 1.00-16.55). Using binary logistic regression analysis, we revealed a relationship between the advanced form of periodontitis and familial history of periodontitis (B= -0.79, OR = 0.45; 95% CI 0.21–0.93). Conclusions Twenty-five individuals in a sample of 156, were positive for A. actinomycetemcomitans . Poor oral hygiene, family history of periodontitis, and irregular attendance to dental apportionments were identified as potential risk factors for severe periodontitis in this investigated sample of Western Australians. Clinical Relevance: A. actinomycetemcomitans is a key pathogen implicated in the pathogenesis of periodontitis. The presence of this pathogen was not documented before in a cohort of Western Australians. We have noted a relatively low presence of A. actinomycetemcomitans in this cohort of Western Australians, which can be compared to some studies in the European population. We also identified some risk factors that can contribute to the severe form of periodontitis in this population. Describing the distribution of A. actinomycetemcomitans and their association with various clinical parameters aid in our understanding of the natural history of periodontal disease in the population of Western Australians.
The objective was to evaluate using unstimulated saliva in detecting Aggregatibacter actinomycetemcomitans and to compare the saliva and subgingival and mucosa membrane occurrence of this periodontal pathogen in patients diagnosed with advanced periodontitis. Patients with advanced forms of periodontitis (n = 220; mean age: 54.03 ± 03 years) at stage III/IV were sampled. Unstimulated saliva, buccal cheek mucosa, and pooled subgingival plaque samples were collected. The identification of A. actinomycetemcomitans was performed using qPCR. A descriptive analysis and Wilcoxon test and analysis of variance were performed. A. actinomycetemcomitans was isolated from 28.18% of the subjects. A total of 660 samples were obtained, 220 from unstimulated saliva, 220 from buccal cheek mucosa surfaces, and 220 from pooled subgingival plaque samples. A. actinomycetemcomitans was isolated from 21.80% of unstimulated saliva, 19.50% of buccal cheek swabs, and 17.70% of subgingival samples. There was a statistically significant difference between the presence of A. actinomycetemcomitans in the unstimulated saliva samples and in the buccal cheek mucosa swab samples and pooled subgingival plaque samples (p < 0.001). These results suggest that in advanced periodontitis, unstimulated saliva is representative of pooled subgingival plaque/buccal cheek mucosa samples and its use is adequate in the oral detection of A. actinomycetemcomitans in a cohort of patients with stage III and IV periodontitis.
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