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A molecular survey of S. mutans and P. gingivalis oral microbial burden in human saliva using relative endpoint polymerase chain reaction (RE-PCR) within the population of a Nevada dental school revealed disparities among minorities.

Davis JE, Freel N, Findley A, Tomlin K, Howard KM, Seran CC, Cruz P, Kingsley K - BMC Oral Health (2012)

Bottom Line: Relative endpoint polymerase chain reaction (RE-PCR) was performed to ascertain the presence and relative microbial burden of these oral pathogens.Nearly one quarter (13/56) or 23.3% of these patients had elevated levels of S. mutans, while (10/56) and 17.8% of these samples were found to have elevated levels of P. gingivalis, - with (90%) of P. gingivalis-positive samples from minority patients (X2=17.921, d.f. = 1; p<0.0001).Oral health knowledge and practices among minority patients may be strongly influenced by other factors, including education and socioeconomic status, suggesting additional research may be needed to accurately determine the most appropriate standards for care and oral health education within this patient population.

View Article: PubMed Central - HTML - PubMed

Affiliation: Orthodontic Residency Program, School of Dental Medicine, University of Nevada, Las Vegas, NV, USA.

ABSTRACT

Background: The University of Nevada, Las Vegas School of Dental Medicine recently opened an orthodontic treatment clinic to address the needs of the racially and ethnically diverse population of Southern Nevada, primarily focusing on the treatment and care of low-income and minority patients. Although orthodontic treatment and therapy has been shown to induce changes in the oral cavity, much of this evidence was collected from traditional White, teenage orthodontic clinic populations. The primary goal of this study was to describe the microbial burden of the cariogenic and periodontal pathogens, Streptococcus mutans and Porphyromonas gingivalis within the UNLV-SDM patient population.

Methods: Representative saliva samples were collected from healthy adult patients for DNA isolation. Relative endpoint polymerase chain reaction (RE-PCR) was performed to ascertain the presence and relative microbial burden of these oral pathogens.

Results: Nearly one quarter (13/56) or 23.3% of these patients had elevated levels of S. mutans, while (10/56) and 17.8% of these samples were found to have elevated levels of P. gingivalis, - with (90%) of P. gingivalis-positive samples from minority patients (X2=17.921, d.f. = 1; p<0.0001).

Conclusions: These findings of elevated P. gingivalis levels, primarily among minority patients, may suggest underlying oral health practices contributing to adverse oral health conditions within this population. Oral health knowledge and practices among minority patients may be strongly influenced by other factors, including education and socioeconomic status, suggesting additional research may be needed to accurately determine the most appropriate standards for care and oral health education within this patient population.

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RE-PCR using DNA from P. gingivalis (PG) standards and saliva samples. A) DNA standards obtained from PG samples containing 5.0 x 103 - 106 CFU/mL established minimum threshold (CT) and saturation (CS) cycles; (high concentration) 5/0 x 106 CFU/mL CT = C15, CS = C35; (low) 5.0 x 136 CFU/mL , CT = C30, CS = 55. B) RE-PCR at C30 (at low concentration CT = C30, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.8507) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 10/56 had elevated PG levels. Plotting the PG-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high concentrations; Very high (n = 4), high risk (n = 4), moderate (n = 2). No significant differences in gender (not shown) between PG-positive and overall sample demographics were noted, however 90% (n = 9/10) of the PG-positive samples came from Minority patients, which was significantly different than in the overall sample (64.9%) (X2 = 17.921, d.f. = 1; p < 0.0001; M = minority, W = white). In addition, the ages of PG-positive patients were not found to significantly different than those of the study sample (p = 0.05).
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Figure 3: RE-PCR using DNA from P. gingivalis (PG) standards and saliva samples. A) DNA standards obtained from PG samples containing 5.0 x 103 - 106 CFU/mL established minimum threshold (CT) and saturation (CS) cycles; (high concentration) 5/0 x 106 CFU/mL CT = C15, CS = C35; (low) 5.0 x 136 CFU/mL , CT = C30, CS = 55. B) RE-PCR at C30 (at low concentration CT = C30, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.8507) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 10/56 had elevated PG levels. Plotting the PG-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high concentrations; Very high (n = 4), high risk (n = 4), moderate (n = 2). No significant differences in gender (not shown) between PG-positive and overall sample demographics were noted, however 90% (n = 9/10) of the PG-positive samples came from Minority patients, which was significantly different than in the overall sample (64.9%) (X2 = 17.921, d.f. = 1; p < 0.0001; M = minority, W = white). In addition, the ages of PG-positive patients were not found to significantly different than those of the study sample (p = 0.05).

Mentions: Standards of genomic DNA extracted from PG samples containing 5.0 x 103 - 106 CFU/mL were used to establish detection threshold and saturation (CT and CS) cycle limits (Figure 3A). For the DNA from samples with the highest CFU/mL concentrations of PG (5.0 x 106 CFU/mL), CT was observed at C15 and CS at C35, similar to the results with the SM standards. CT was established at approximately 20, 25 and 30 for each successful sample dilution (105, 104, and 103 CFU/mL, respectively), with CS at correspondingly higher cycles (~C45 – C55). The previously established GAPDH EP cycle C30 was therefore found to be at CT for DNA samples with CFU/mL concentrations in the lowest category (CT = C30), and above the CT for DNA samples from the higher categories (CT = C15 – 25), as well as being below the upper limit for the highest concentration, CS = C35). In addition, the ages of PG-positive patients were not found to be significantly different than those of the study sample (p = 0.05).


A molecular survey of S. mutans and P. gingivalis oral microbial burden in human saliva using relative endpoint polymerase chain reaction (RE-PCR) within the population of a Nevada dental school revealed disparities among minorities.

Davis JE, Freel N, Findley A, Tomlin K, Howard KM, Seran CC, Cruz P, Kingsley K - BMC Oral Health (2012)

RE-PCR using DNA from P. gingivalis (PG) standards and saliva samples. A) DNA standards obtained from PG samples containing 5.0 x 103 - 106 CFU/mL established minimum threshold (CT) and saturation (CS) cycles; (high concentration) 5/0 x 106 CFU/mL CT = C15, CS = C35; (low) 5.0 x 136 CFU/mL , CT = C30, CS = 55. B) RE-PCR at C30 (at low concentration CT = C30, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.8507) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 10/56 had elevated PG levels. Plotting the PG-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high concentrations; Very high (n = 4), high risk (n = 4), moderate (n = 2). No significant differences in gender (not shown) between PG-positive and overall sample demographics were noted, however 90% (n = 9/10) of the PG-positive samples came from Minority patients, which was significantly different than in the overall sample (64.9%) (X2 = 17.921, d.f. = 1; p < 0.0001; M = minority, W = white). In addition, the ages of PG-positive patients were not found to significantly different than those of the study sample (p = 0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 3: RE-PCR using DNA from P. gingivalis (PG) standards and saliva samples. A) DNA standards obtained from PG samples containing 5.0 x 103 - 106 CFU/mL established minimum threshold (CT) and saturation (CS) cycles; (high concentration) 5/0 x 106 CFU/mL CT = C15, CS = C35; (low) 5.0 x 136 CFU/mL , CT = C30, CS = 55. B) RE-PCR at C30 (at low concentration CT = C30, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.8507) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 10/56 had elevated PG levels. Plotting the PG-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high concentrations; Very high (n = 4), high risk (n = 4), moderate (n = 2). No significant differences in gender (not shown) between PG-positive and overall sample demographics were noted, however 90% (n = 9/10) of the PG-positive samples came from Minority patients, which was significantly different than in the overall sample (64.9%) (X2 = 17.921, d.f. = 1; p < 0.0001; M = minority, W = white). In addition, the ages of PG-positive patients were not found to significantly different than those of the study sample (p = 0.05).
Mentions: Standards of genomic DNA extracted from PG samples containing 5.0 x 103 - 106 CFU/mL were used to establish detection threshold and saturation (CT and CS) cycle limits (Figure 3A). For the DNA from samples with the highest CFU/mL concentrations of PG (5.0 x 106 CFU/mL), CT was observed at C15 and CS at C35, similar to the results with the SM standards. CT was established at approximately 20, 25 and 30 for each successful sample dilution (105, 104, and 103 CFU/mL, respectively), with CS at correspondingly higher cycles (~C45 – C55). The previously established GAPDH EP cycle C30 was therefore found to be at CT for DNA samples with CFU/mL concentrations in the lowest category (CT = C30), and above the CT for DNA samples from the higher categories (CT = C15 – 25), as well as being below the upper limit for the highest concentration, CS = C35). In addition, the ages of PG-positive patients were not found to be significantly different than those of the study sample (p = 0.05).

Bottom Line: Relative endpoint polymerase chain reaction (RE-PCR) was performed to ascertain the presence and relative microbial burden of these oral pathogens.Nearly one quarter (13/56) or 23.3% of these patients had elevated levels of S. mutans, while (10/56) and 17.8% of these samples were found to have elevated levels of P. gingivalis, - with (90%) of P. gingivalis-positive samples from minority patients (X2=17.921, d.f. = 1; p<0.0001).Oral health knowledge and practices among minority patients may be strongly influenced by other factors, including education and socioeconomic status, suggesting additional research may be needed to accurately determine the most appropriate standards for care and oral health education within this patient population.

View Article: PubMed Central - HTML - PubMed

Affiliation: Orthodontic Residency Program, School of Dental Medicine, University of Nevada, Las Vegas, NV, USA.

ABSTRACT

Background: The University of Nevada, Las Vegas School of Dental Medicine recently opened an orthodontic treatment clinic to address the needs of the racially and ethnically diverse population of Southern Nevada, primarily focusing on the treatment and care of low-income and minority patients. Although orthodontic treatment and therapy has been shown to induce changes in the oral cavity, much of this evidence was collected from traditional White, teenage orthodontic clinic populations. The primary goal of this study was to describe the microbial burden of the cariogenic and periodontal pathogens, Streptococcus mutans and Porphyromonas gingivalis within the UNLV-SDM patient population.

Methods: Representative saliva samples were collected from healthy adult patients for DNA isolation. Relative endpoint polymerase chain reaction (RE-PCR) was performed to ascertain the presence and relative microbial burden of these oral pathogens.

Results: Nearly one quarter (13/56) or 23.3% of these patients had elevated levels of S. mutans, while (10/56) and 17.8% of these samples were found to have elevated levels of P. gingivalis, - with (90%) of P. gingivalis-positive samples from minority patients (X2=17.921, d.f. = 1; p<0.0001).

Conclusions: These findings of elevated P. gingivalis levels, primarily among minority patients, may suggest underlying oral health practices contributing to adverse oral health conditions within this population. Oral health knowledge and practices among minority patients may be strongly influenced by other factors, including education and socioeconomic status, suggesting additional research may be needed to accurately determine the most appropriate standards for care and oral health education within this patient population.

Show MeSH
Related in: MedlinePlus