Use of synthetic oligonucleotide DNA probes for the identification of Bacteroides gingivalis

Abstract

Six different oligonucleotide probes complementary to the hypervariable regions of 16S rRNA of Bacteroides gingivalis were tested for specificity and sensitivity against 77 field strains of B. gingivalis and 105 strains of 12 other Bacteroides species. The data demonstrated that these probes were very specific (range, 0.85 to 1.00) and sensitive (1.00). Some limited cross-reactions with other Bacteroides species were observed. Four of these probes should be useful for rapid detection and identification of B. gingivalis.

Intra- and inter-individual comparison of Porphyromonas gingivalis genotypes

“…Chromosomal DNA from the bacterial cells cultured on supplemented Brucella blood agar plates (1-2 plates per strain) was extracted by the method of Moncla et al [1]. DNA (2-3 ~g) isolated from all 31 strains was digested to completion with restriction endonucleases PstI, ClaI and BglI (Boehringer Mannheim, FRG) according to the manufacturer's instructions.…”

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Genetic analysis of 31 clinical strains of Porphyromonas gingivalis isolated from nine subjects, 2-6 strains per subject, was performed by Southern hybridization. Chromosomal DNA was extracted by the method of Moncla et al [1] and digested to completion with restriction endonucleases Pst I, ClaI and BglI. The DNA fragments were separated electrophoretically on agarose gels, transferred to nylon membranes and hybridized to the non-radioactively labelled plasmid pKK 3535 which contains the rrnB ribosomal RNA operon of the Escherichia coli chromosome.

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Microbial changes in patients with acute periodontal abscess after treatment detected by PadoTest

“…The two methods had an agreement of around 80% for T. forsythia, P. gingivalis, and T. denticola and 70% for A. actinomycetemcomitans (Geinoz et al, 1997). Studies reporting the specificity of oligonucleotide probe method showed that all oligonucleotide probes had species specificity by cross-reaction checks, except few cases between P. gingivalis and related species (Dix et al, 1990;Moncla et al, 1990). The PadoTest 4.5 has been used to evaluate the changes in microbial load in a few studies (Kamma and Baehni, 2003;Mombelli et al, 2005).…”

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

“…At present, the status and progress of periodontal disease is monitored by clinical parameters such as gingival inflammation, bleeding index, depth of periodontal pockets and loss of alveolar bone. Considering the microbiologybased definition of activity, screening for the 116 presence and number of potential bacterial pathogens on a routine basis could be a helpful supplement to the diagnosis of periodontitis, but inherent drawbacks such as low specificity, time or cost effectiveness of conventional microbiological techniques (immunofluorescence, microscopy and culture methods) have prevented the routine use of these techniques [15][16][17][18].…”

“…The use of synthetic and species-specific DNA probes directed against 16S rRNA sequences of periodontopathogenic bacteria has been considered as a promising method for routine screening of subgingival plaque and exudate of patients with marginal periodontitis [17][18][19]. To achieve a high sensitivity, probes are commonly labelled radioactively with 32p.…”