Non-radioactively labelled DNA probes for the detection of periodontopathogenic Prevotella and Porphyromonas species

Abstract

Digoxigenin-labelled synthetic DNA probes directed against the 16S rRNA were used for the direct detection of the periodontopathogenic bacteria Prevotella intermedia and Porphyromonas gingivalis in subgingival plaque by applying a DNA-RNA dot-blot hybridization procedure. The test was evaluated with 134 plaque samples from 26 patients with adult periodontitis or rapidly progressive periodontitis. The lower limit of detection was 10(4)-10(5) bacteria/specimen. A semiquantitative assessment of the two species in each sample and in the corresponding periodontal site was achieved by this technique. It is possible to examine 80-90 samples within two days with low material costs.

Microbiology of periodontal disease in children and young adults

“…Lopez et al (90) isolated P. gingivalis from 70% of localized early-onset periodontitis patients and 67% of sites, but a number of other organisms were more prevalent. This same group, in a later study (91), detected P. gingivalis in 17 of 18 localized early-onset periodontitis patients, and they suggested that localized early-onset periodontitis is more frequently associated with P. gingivalis (and Prevotella intermedia) 36 given the lower prevalence of A. actinomycetemcomitans. Moore et al (106), in their studies of localized early-onset periodontitis patients, reported that 6% of periodontal sites were positive for P. gingivalis and the organism's proportion of the flora was 12-fold greater in diseased sites compared with healthy sites.…”

“…P. gingivalis has also been implicated in generalized early‐onset periodontitis in a number of studies (3, 36, 73–75, 85, 88, 90, 91, 97, 104, 118, 141, 174) (Table 3) and is often the predominant organism in the disease (2, 73, 74, 90, 118, 174). Wilson et al (182) found that 8% to 16% of the cultivable microflora was P. gingivalis and suggested that P. gingivalis was involved in causing generalized early‐onset periodotitis.…”

“…Williams et al (182) did not detect P. gingivalis in their study of generalized early‐onset periodontitis. P. gingivalis has also been frequently isolated with P. intermedia (2, 36, 97, 174), A. actinomycetemcomitans (73, 85, 97), F. nucleatum (118) or a combination of these including Bacteroides forsythus (74, 75, 141).…”

“…P. intermedia has also been associated with generalized early‐onset periodontitis. It has been found at a higher prevalence and in higher numbers (3, 36, 73, 74, 88, 90, 91, 97, 104, 174) and it can be the predominant organism (141, 182). In two studies by Lopez et al (90, 91) they reported that P. intermedia was found in all generalized early‐onset periodontitis patients and sites in one group and in 40% of similar patients and sites in another group.…”

Oligonucleotide probes to the 16s ribosomal RNA: implications of sequence homology and secondary structure with particular reference to the oral species Prevotetla intermedia and Prevotetla nigrescens

“…Several attempts have been made to find reliable nucleic acid probes to distinguish P. intermedia from P. nigrescens. The most widely reported are oligonucleotide signatures derived from the 16s rRNA molecule which have also been adopted for commercial and clinical use (see eg French et al, 1986;Dix et al, 1990;Conrads and Brauner, 1993). Comparative analysis of five sequences of the small-subunit structure of the ribosomal RNA from both species indicated that they are phylogenetically closely related (92-95%).…”