Comparison of DNA Probe and ELISA Microbial Analysis Methods and Their Association With Adult Periodontitis

Abstract

The purposes of this study were two‐fold: to compare the DNA probe and enzyme linked immunosorbent assay (ELISA) microbial identification tests and correlate the levels of microorganisms with adult periodontitis. A single plaque sample was taken from each of 2 sites in 52 patients. Twelve of these patients were also sampled during and after treatment. The experimental site had clinical indicators of disease (bleeding on probing, probing and attachment loss of ≥ 6 mm) and the contralateral site (control) was clinically healthy. A total of 176 plaque samples were collected, divided, processed, and sent for both types of quantitative microbial analyses. All of these samples were used to compare the DNA probe and ELISA methods while only the initial 104 pretreatment sites were used to correlate microorganisms/method with clinical indicators of adult periodontitis. DNA probes were used to assay for A. actinomycetemcomitans, P. gingivalis, P. intermedia, E. corrodens, F. nucleatum, T. denticola, and C. rectus. An ELISA utilizing monoclonal antibodies was used to assay for P. gingivalis, E. corrodens, T. denticola, and C. rectus. Comparison of the two methods revealed that the ELISA test identified P. gingivalis and C. rectus significantly more often than the DNA probe method and that T. denticola was detected more frequently with the DNA probe. The sensitivities and specificities varied widely among organisms and by test. P. gingivalis, as identified by ELISA, had the highest degree of sensitivity and specificity (0.90 and 0.82 respectively) to clinical indicators of adult periodontitis. J Periodontol 1994;65:576–582.

Comparison of 16S rDNA-based PCR and checkerboard DNA–DNA hybridisation for detection of selected endodontic pathogens

“…Cross-reactivity for the whole DNA probe may explain the PCR-negative/checkerboard-positive results [30]. Although whole genomic probes are more likely to cross-react with non-target bacteria due to the presence of homologous sequences between different bacterial species [29], the results of this study suggested that this was not a common occurrence.…”

Periodontal Disease as a Specific, albeit Chronic, Infection: Diagnosis and Treatment

“…The sensitivity ranged from 0.17 for A. actinomycetemcomitans to 0.86 for B. forsythus and Streptococcus sanguis, and the specificity ranged from 0.17 for P. intermedia to 1.0 for C. rectus. These ranges indicate the typical finding with DNA probes, namely, that they do not correlate well with culture or serological data, usually being positive when the culture or serological findings are negative, i.e., many false positives (142,158,172,179,191,195,221,255,345). The panel of 18 probes developed by Papapanou showed that only B. forsythus, P. gingivalis, T. denticola, and C. rectus (Wolinella recta) were associated with periodontal disease in 148 Chinese subjects who had never received any periodontal treatment (220).…”

“…A whole-genome probe to B. forsythus showed no detectable reactivity with 75 strains representing 14 oral species (172). This limited evaluation means that some crossreactions with untested plaque species probably exist, and could contribute to the large number of false-positive results found when DNA probes are compared to culture results obtained on the same plaque samples (15,135,142,158,172,191,195,196,221,235,251,255,296,297,315,345; K. Backman, Letter, J. Periodontol. 66:536-537, 1995).…”

A Case‐Control Study on the Association of Human Leukocyte Antigen‐A*9 and ‐B*15 Alleles With Generalized Aggressive Periodontitis in an Indian Population

“…Aggressive periodontitis has a multifactorial etiology, and the characteristic tissue destruction is mediated mainly by the aberrant immune response of the host to periodontopathic bacteria. The role of oral microflora in the etiology of various inflammatory periodontal diseases has been well established, and specificity may vary between bacterial etiologies and different forms of periodontal disease 4‐8 . However, environmental factors alone cannot explain the occurrence of aggressive periodontitis.…”