Identification of Bacteroides forsythus in Subgingival Dental Plaque with the Aid of a Rapid PCR Method

Abstract

Bacteroides forsythus has been shown to be prevalent among patients with periodontitis. Conventional microbiological methods used to identify this bacterium, however, are laborious and time-consuming and are therefore not well-suited for screening purposes. We have developed a polymerase chain-reaction (PCR) method which is rapid, specific, and simple to perform and does not require other sample pre-treatment except a brief centrifugation. This method was applied to the detection of B. forsythus in subgingival plaque of 58 periodontitis patients. When compared with the results of conventional culturing, the PCR method always confirmed the culture-positive results, while none of the PCR negative samples was shown to be culture-positive. The PCR method appeared to give more than double the number of samples positive for B. forsythus than culturing (89.7% vs. 37.9%). The analysis requires less than 4 hrs to perform, and is specific only to B. forsythus and sensitive enough to detect fewer than 5 bacteria.

Keywords

Bacteroides forsythus human periodontal disease polymerase chain reaction.

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References

Braham PH , Moncla BJ (1992). Rapid presumptive identification and further characterization of Bacteroides forsythus. J Clin Microbiol 30:649-654.

Conrads G. , Brauner A. (1995). Detection of periodontopathic bacteria by DNA probes and therapeutic consequences. In: Medical and dental aspects of anaerobes. Duerden BI , Wade WG , Brazier JS , Eley A , Wren B , Hudson MJ , editors. Northwood: Science Reviews, pp. 63-71.

Cross DL , Smith Glf (1995). DNA probes for periodontal pathogens in HIV patients . In: Medical and dental aspects of anaerobes. Duerden BI , Wade WG , Brazier JS , Eley A , Wren B , Hudson MJ , editors. Northwood: Science Reviews, pp. 387-402.

Declaration of Helsinki (1975). Guidelines for doctors using humans in biomedical research. Approved by the 18th World Medical Association Assembly in Helsinki, Finland, and revised at the 29th World Medical Association Assembly in Tokyo, Japan.

Gersdorf H. , Meissner A. , Pelz K. , Krekeler G. , Gobel UB (1993). Identification of Bacteroides forsythus in subgingival plaque from patients with advanced periodontitis. J Clin Microbiol 31:941-946.

Gmür R. , Guggenheim B. (1990). Monoclonal antibodies for the detection of "periodontopathic" bacteria. Arch Oral Biol 35(Suppl):145S-151S.

Ishihara K. , Naito Y. , Kato T. , Takazoe I. , Okuda K. , Eguchi T. , et al. (1992). A sensitive enzymatic method (SK-013) for detection and quantification of specific periodontopathogens. J Periodont Res 27:81-85.

Listgarten MA , Lai CH , Young V. (1993). Subgingival microbial samples from patients with refractory periodontitis. J Periodontol 64:155-161.

Loesche WJ , Bretz WA , Kerschensteiner D. , Stoll J. , Socransky SS , Hujoel P. , et al. (1990). Development of a diagnostic test for anaerobic periodontal infections based on plaque hydrolysis of benzoyl-DL-arginine-naphthylamide . J Clin Microbiol 28:1551-1559.

10.

Loesche WJ , Lopatin DE , Stoll J. , van Poperin N. , Hujoel P. (1992a). Comparison of various detection methods for periodontopathic bacteria: can culture be considered the primary reference standard? J Clin Microbiol 30:418-426.

11.

Loesche WJ , Lopatin DE , Giordano J. , Alcoforado G. , Hujoel P. (1992b). Comparison of the benzoyl-DL-arginine-naphthylamide (BANA) test, DNA probes, and immunological reagents for ability to detect anaerobic periodontal infections due to Porphyromonas gingivalis, Treponema denticola, and Bacteroides forsythus. J Clin Microbiol 30:427-433.

12.

Mombelli A. , Lang NP , Burgin WB , Gusberti FA (1990). Microbial changes associated with the development of puberty gingivitis. J Periodontol 25:331-338.

13.

Moncla BJ , Motley ST , Braham P. , Ewing L. , Adams TH , Vermeulen NM (1991). Use of synthetic oligonucleotide DNA probes for identification and direct detection of Bacteroides forsythus in plaque samples . J Clin Microbiol 29:2158-2162.

14.

Moller Ajr (1966). Microbiological examination of root canals and periapical tissues of human teeth. Methodological studies. Odontol Tidskr 74(Suppl):1-380.

15.

Seida K. , Saito A. , Yamada S. , Ishihara K. , Naito Y. , Okuda K. (1992). A sensitive enzymatic method (SK-013) for detection of Treponema denticola, Porphyromonas gingivalis and Bacteroides forsythus in subgingival plaque samples. J Periodont Res 27:86-91.

16.

Tanner A. , Bouldin HD , Maiden MF (1989). Newly delineated periodontal pathogens with special reference to Selenomonas species. Infection 17:182-187.

17.

Wahlfors J. , Meurman JH , Toskala J. , Korhonen A. , Alakuijala P. , Janatuinen E. , et al. (1995a). Development of a rapid PCR method for identification of Helicobacter pylori in dental plaque and gastric biopsy specimens. Eur J Clin Microbiol Infect Dis 14:780-786.

18.

Wahlfors J. , Meurman JH , Vaisanen P. , Alakuijala P. , Korhonen A. , Torkko H. , et al. (1995b). Simultaneous detection of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis by a rapid PCR method. J Dent Res 74:1796-1801.

19.

Wilson MI , Wade G. , Weightinan AJ (1995). 16S ribosomal sequencing: analysis of the oral microflora. In: Medical and dental aspects of anaerobes. Duerden BI , Wade WG , Biazici JS , Eley A , Wren B , Hudson MI , editors. Northwood: Science Reviews, p. 73.