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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 S. mutans (SM) standards and saliva samples. A) DNA standards obtained from SM 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 = C40; (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 = C40) revealed strong, positive correlations (R2 = 0.9645) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 13/56 had elevated SM levels. Plotting the SM-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high caries risk; Very high caries risk (n = 3), high risk (n = 5), moderate risk (n = 5). No significant differences in gender (not shown) or race/ethnicity between SM-positive and overall sample demographics were noted (M = minority, W = white). No statistically significant differences were found among the ages of SM-positive samples and those of the study sample (p = 0.2798).
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Figure 2: RE-PCR using DNA from S. mutans (SM) standards and saliva samples. A) DNA standards obtained from SM 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 = C40; (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 = C40) revealed strong, positive correlations (R2 = 0.9645) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 13/56 had elevated SM levels. Plotting the SM-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high caries risk; Very high caries risk (n = 3), high risk (n = 5), moderate risk (n = 5). No significant differences in gender (not shown) or race/ethnicity between SM-positive and overall sample demographics were noted (M = minority, W = white). No statistically significant differences were found among the ages of SM-positive samples and those of the study sample (p = 0.2798).

Mentions: Standards of genomic DNA extracted from SM samples containing 5.0 x 103 - 106 CFU/mL were used to establish detection threshold and saturation (CT and CS) cycle limits (Figure 2A). For the DNA samples with the highest CFU/mL concentration (5.0 x 106 CFU/mL), CT was observed at C15 and CS at C40. CT was established at approximately 20, 25 and 30 for each successful sample dilution (105, 104, and103 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 for average caries risk (CT = C30), and above the CT for DNA samples with moderate, high, or very high risk (CT = C15 - C25), as well as being below the upper limit for the highest concentration, CS = C40.


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 S. mutans (SM) standards and saliva samples. A) DNA standards obtained from SM 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 = C40; (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 = C40) revealed strong, positive correlations (R2 = 0.9645) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 13/56 had elevated SM levels. Plotting the SM-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high caries risk; Very high caries risk (n = 3), high risk (n = 5), moderate risk (n = 5). No significant differences in gender (not shown) or race/ethnicity between SM-positive and overall sample demographics were noted (M = minority, W = white). No statistically significant differences were found among the ages of SM-positive samples and those of the study sample (p = 0.2798).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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Figure 2: RE-PCR using DNA from S. mutans (SM) standards and saliva samples. A) DNA standards obtained from SM 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 = C40; (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 = C40) revealed strong, positive correlations (R2 = 0.9645) between signal band intensity (SBI) and CFU/mL. C) RE-PCR using DNA extractions from all saliva samples revealed (n = 13/56 had elevated SM levels. Plotting the SM-positive sample SBI (*) with the DNA standards revealed samples with moderate to very high caries risk; Very high caries risk (n = 3), high risk (n = 5), moderate risk (n = 5). No significant differences in gender (not shown) or race/ethnicity between SM-positive and overall sample demographics were noted (M = minority, W = white). No statistically significant differences were found among the ages of SM-positive samples and those of the study sample (p = 0.2798).
Mentions: Standards of genomic DNA extracted from SM samples containing 5.0 x 103 - 106 CFU/mL were used to establish detection threshold and saturation (CT and CS) cycle limits (Figure 2A). For the DNA samples with the highest CFU/mL concentration (5.0 x 106 CFU/mL), CT was observed at C15 and CS at C40. CT was established at approximately 20, 25 and 30 for each successful sample dilution (105, 104, and103 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 for average caries risk (CT = C30), and above the CT for DNA samples with moderate, high, or very high risk (CT = C15 - C25), as well as being below the upper limit for the highest concentration, CS = C40.

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