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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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Related in: MedlinePlus

RE-PCR using DNA from HGF-1 cells (standards) and saliva samples. A) DNA standards obtained from HGF-1 cells (0.5 – 2.5 x 106 cells/mL) established minimum threshold (CT) and saturation (CS) cycles; (high cell concentration) 2.5 x 106 cells/mL CT = C10, CS = C35; (low cell concentration) 0.5 x 106 cells/mL , CT = C20, CS = C45. B) RE-PCR at C30 (above low concentration CT = C20, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.9918) between signal band intensity (SBI) and cell concentration. C) RE-PCR using DNA extractions from all saliva samples produced bands with increasing SBI; two representative saliva samples with low (0.8 – 1.2 x 106 cells/mL), mid (1.6 – 1.9 x 106 cells/mL) and high (2.1 – 2.4 x 106 cells/mL) cell concentrations are shown. Plotting the sample SBI (*) with the DNA standards revealed near perfect alignment.
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Figure 1: RE-PCR using DNA from HGF-1 cells (standards) and saliva samples. A) DNA standards obtained from HGF-1 cells (0.5 – 2.5 x 106 cells/mL) established minimum threshold (CT) and saturation (CS) cycles; (high cell concentration) 2.5 x 106 cells/mL CT = C10, CS = C35; (low cell concentration) 0.5 x 106 cells/mL , CT = C20, CS = C45. B) RE-PCR at C30 (above low concentration CT = C20, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.9918) between signal band intensity (SBI) and cell concentration. C) RE-PCR using DNA extractions from all saliva samples produced bands with increasing SBI; two representative saliva samples with low (0.8 – 1.2 x 106 cells/mL), mid (1.6 – 1.9 x 106 cells/mL) and high (2.1 – 2.4 x 106 cells/mL) cell concentrations are shown. Plotting the sample SBI (*) with the DNA standards revealed near perfect alignment.

Mentions: DNA standards obtained from standardized control cells, human gingival fibroblasts (0.5 – 2.5 x 106 cells/mL), approximating the range of cell concentrations observed in the saliva samples (0.8 – 2.4 x 106 cells/mL) were used to establish the minimum threshold (CT) and saturation (CS) cycles required for calibration and concentration comparisons using relative endpoint PCR (Figure 1A). For the DNA standard with the highest cell concentration, 2.5 x 106 cells/mL, GAPDH signal detection above background or CT required a minimum of ten cycles (C10), with saturation or CS observed at C35. CT was established at C20 using the DNA standard from the lowest cell concentration, 0.5 x 106 cells/mL, with CS observed at C45. Based upon these data, RE-PCR was performed at C30, an EP cycle above the lower cell concentration CT = C20, but below the higher cell concentration CS = C35.


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 HGF-1 cells (standards) and saliva samples. A) DNA standards obtained from HGF-1 cells (0.5 – 2.5 x 106 cells/mL) established minimum threshold (CT) and saturation (CS) cycles; (high cell concentration) 2.5 x 106 cells/mL CT = C10, CS = C35; (low cell concentration) 0.5 x 106 cells/mL , CT = C20, CS = C45. B) RE-PCR at C30 (above low concentration CT = C20, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.9918) between signal band intensity (SBI) and cell concentration. C) RE-PCR using DNA extractions from all saliva samples produced bands with increasing SBI; two representative saliva samples with low (0.8 – 1.2 x 106 cells/mL), mid (1.6 – 1.9 x 106 cells/mL) and high (2.1 – 2.4 x 106 cells/mL) cell concentrations are shown. Plotting the sample SBI (*) with the DNA standards revealed near perfect alignment.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3473244&req=5

Figure 1: RE-PCR using DNA from HGF-1 cells (standards) and saliva samples. A) DNA standards obtained from HGF-1 cells (0.5 – 2.5 x 106 cells/mL) established minimum threshold (CT) and saturation (CS) cycles; (high cell concentration) 2.5 x 106 cells/mL CT = C10, CS = C35; (low cell concentration) 0.5 x 106 cells/mL , CT = C20, CS = C45. B) RE-PCR at C30 (above low concentration CT = C20, below high concentration CS = C35) revealed strong, positive correlations (R2 = 0.9918) between signal band intensity (SBI) and cell concentration. C) RE-PCR using DNA extractions from all saliva samples produced bands with increasing SBI; two representative saliva samples with low (0.8 – 1.2 x 106 cells/mL), mid (1.6 – 1.9 x 106 cells/mL) and high (2.1 – 2.4 x 106 cells/mL) cell concentrations are shown. Plotting the sample SBI (*) with the DNA standards revealed near perfect alignment.
Mentions: DNA standards obtained from standardized control cells, human gingival fibroblasts (0.5 – 2.5 x 106 cells/mL), approximating the range of cell concentrations observed in the saliva samples (0.8 – 2.4 x 106 cells/mL) were used to establish the minimum threshold (CT) and saturation (CS) cycles required for calibration and concentration comparisons using relative endpoint PCR (Figure 1A). For the DNA standard with the highest cell concentration, 2.5 x 106 cells/mL, GAPDH signal detection above background or CT required a minimum of ten cycles (C10), with saturation or CS observed at C35. CT was established at C20 using the DNA standard from the lowest cell concentration, 0.5 x 106 cells/mL, with CS observed at C45. Based upon these data, RE-PCR was performed at C30, an EP cycle above the lower cell concentration CT = C20, but below the higher cell concentration CS = C35.

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