1887

Abstract

Peptostreptococci are gram-positive, strictly anaerobic bacteria which, although regarded as members of the commensal human microflora, are also frequently isolated from sites of clinical infection. The study of this diverse group of opportunist pathogens has been hindered by an inadequate taxonomy and the lack of a valid identification scheme. Recent re-classification of the Peptostreptococcus family into five distinct genus groups has helped to clarify the situation. However, this has been on the basis of 16S rRNA sequence determinations, which are both time-consuming and expensive. The aim of the present study was to evaluate the use of PCR-amplified ribosomal DNA spacer polymorphisms for the rapid differentiation of the currently recognised taxa within the group of anaerobic gram-positive cocci. A collection comprising 19 reference strains with representatives of each of the 15 species, two close relatives and two of the well-characterised groups, together with 38 test strains was studied. All strains were identified to species group level by phenotypic means. Amplification of the 16S–23S intergenic spacer region (ISR) with universal primers produced distinct banding patterns for all the 19 reference strains and the patterns could be differentiated easily visually. However, of the 38 test strains, less than half could be speciated from ISR analysis alone. Only five groups produced correlating banding patterns for all members tested (, , , and ). For other species, either the type strain differed significantly from other species members (e.g., ) or there appeared to be considerable intra-species variation (e.g., ). Partial 16S rRNA gene sequences for the ‘’ and ‘βGAL’ groups showed that both are most closely related to the group. This work highlights the heterogeneous nature of a number of species and hence the need for still further revision of the taxonomy of this important group of pathogens.

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2002-11-01
2019-11-18
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References

  1. Rodloff AC, Hillier SL, Moncla BJ. Peptostrepotococcus, Propionibacterium, Lactobacillus, Actinomyces and other non-sporing anaerobic gram-positive bacteria. In: Murray PR, Baron EJ, Pfaller MA, Tenover FC, Yolken (eds) Manual of clinical microbiology, 7th edn. Washington, DC, ASM Press. 1999:; 672–689.
  2. Brook I. Recovery of anaerobic bacteria from clinical specimens in 12 years at two military hospitals. J Clin Microbiol 1988;; 26: 1181–1188.
    [Google Scholar]
  3. Holland JW, Hill EO, Altemeier WA. Numbers and types of anaerobic bacteria isolated from clinical specimens since 1960. J Clin Microbiol 1977;; 5: 20–25.
    [Google Scholar]
  4. Murdoch DA, Mitchelmore IJ, Tabaqchali S. The clinical importance of gram-positive anaerobic cocci isolated at St Bartholomew's Hospital, London, in 1987. J Med Microbiol 1994;; 41: 36–44.[CrossRef]
    [Google Scholar]
  5. Wren MWD, Baldwin AWF, Eldon CP, Sanderson PJ. The anaerobic culture of clinical specimens: a 14-month study. J Med Microbiol 1977;; 10: 49–61.[CrossRef]
    [Google Scholar]
  6. Murdoch DA. Gram-positive anaerobic cocci. Clin Microbiol Rev 1998;; 11: 81–120.
    [Google Scholar]
  7. Collins MD, Lawson PA, Willems A.et al. The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. Int J Syst Bacteriol 1994;; 44: 812–826.[CrossRef]
    [Google Scholar]
  8. Conrads G, Soffner J, Pelz K, Mutters R. Taxonomic update and clinical significance of species within the genus Peptostreptococcus. Clin Infect Dis 1997;; 25 Suppl 2: S94–S97.[CrossRef]
    [Google Scholar]
  9. Li N, Hashimoto Y, Ezaki T. Determination of 16S ribosomal RNA sequences of all members of the genus Peptostreptococcus and their phylogenetic position. FEMS Microbiol Lett 1994;; 116: 1–6.[CrossRef]
    [Google Scholar]
  10. Murdoch, DA, Magee JT. A numerical taxonomic study of the gram-positive anaerobic cocci. J Med Microbiol 1995;; 43: 148–155.[CrossRef]
    [Google Scholar]
  11. Murdoch DA, Shah HN, Gharbia SE, Rajendram D. Proposal to restrict the genus Peptostreptococcus (Kluyver and van Niel 1936) to Peptostreptococcus anaerobius. Anaerobe 2000;; 6: 257–260.[CrossRef]
    [Google Scholar]
  12. Murdoch DA, Collins MD, Willems A, Hardie JM, Young KA, Magee JT. Description of three new species of the genus Peptostreptococcus from human clinical specimens: Peptostreptococcus harei sp. nov., Peptostreptococcus ivorii sp. nov., and Peptostreptococcus octavius sp. nov. Int J Sys Bacteriol 1997;; 47: 781–787.[CrossRef]
    [Google Scholar]
  13. Murdoch DA, Shah HN. Reclassification of Peptostreptococcus magnus (Prevot 1933) Holdemann and Moore 1972 as Finegoldia magna comb. nov. and Peptostreptococcus micros (Prevot 1933) Smith 1957 as Micromonas micros comb. nov. Anaerobe 1999;; 5: 555–559.[CrossRef]
    [Google Scholar]
  14. Ezaki T, Kawamura Y, Li N, Li Z-Y, Zhao L, Shu S. Proposal of the genera Anaerococcus gen. nov., Peptoniphilus gen. nov. and Gallicola gen. nov. for members of the genus Peptostreptococcus. Int J Syst Evol Microbiol 2001;; 51: 1521–1528.
    [Google Scholar]
  15. Barry T, Colleran G, Glennon M, Duncan LK, Gannon F. The 16S/23S ribosomal spacer region as a target for DNA probes to identify eubacteria. PCR Methods Appl 1991;; 1: 51–56.[CrossRef]
    [Google Scholar]
  16. Jensen MA, Webster JA, Strauss N. Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms. Appl Environ Microbiol 1993;; 59: 945–952.
    [Google Scholar]
  17. Kostman JR, Alden MB, Mair M, Edlind TE, Lipuma JJ, Stull TL. A universal approach to bacterial molecular epidemiology by polymerase chain reaction ribotyping. J Infect Dis 1995;; 171: 204–208.[CrossRef]
    [Google Scholar]
  18. Tyrrell GJ, Bethune RN, Willey B, Low DE. Species identification of enterococci via intergenic ribosomal PCR. J Clin Microbiol 1997;; 35: 1054–1060.
    [Google Scholar]
  19. Ng L-K, Dillon J-A. Molecular fingerprinting of isolates of the genus Peptostreptococcus using rRNA genes from Escherichia coli and P. anaerobius. J Gen Microbiol 1991;; 137: 1323–1331.[CrossRef]
    [Google Scholar]
  20. Holdeman LV, Moore WEC, Cato EP (eds). Anaerobic laboratory manual, 4th edn. Blacksburg, VA, Virginia Polytechnic Institute and State University. 1977.
  21. Phillips KD. A simple and sensitive technique for determining the fermentation reactions of non-sporing anaerobes. J Appl Bacteriol 1976;; 41: 325–328.[CrossRef]
    [Google Scholar]
  22. Willis AT, Phillips KD. Anaerobic infections, 2nd edn. London, HMSO. 1983.
  23. de Lamballerie X, Zandotti C, Vignoli C, Bollet C, de Micco P. A one-step microbial DNA extraction method using ‘‘Chelex-100’’ suitable for gene amplification. Res Microbiol 1992;; 143: 785–790.[CrossRef]
    [Google Scholar]
  24. Marchesi JR, Sato T, Weightman AJ.et al. Design and evaluation of useful bacterial-specific PCR primers that amplify genes coding for bacterial 16S rRNA genes. Appl Environ Microbiol 1998;; 64: 795–799.
    [Google Scholar]
  25. Stoesser G, Baker W, van den Broek A.et al. The EMBL nucleotide sequence database. Nucleic Acids Res 2001;; 29: 17–21.[CrossRef]
    [Google Scholar]
  26. Pearson WR, Lipman DJ. Improved tools for biological sequence analysis. PNAS 1988;; 85: 2444–2448.[CrossRef]
    [Google Scholar]
  27. Pearson WR. Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol 1990;; 183: 63–98.
    [Google Scholar]
  28. Maidak BL, Cole JR, Lilburn TG.et al. The RDP-II (Ribosomal Database Project). Nucleic Acids Res 2001;; 29: 173–174.[CrossRef]
    [Google Scholar]
  29. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994;; 22: 4673–4680.[CrossRef]
    [Google Scholar]
  30. Jukes TH, Cantor CR. Evolution of protein molecules. In: Munro HN (ed) Mammalian protein metabolism, vol 3.New York, Academic Press. 1969:; 21–132.
  31. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;; 4: 406–425.
    [Google Scholar]
  32. Van de Peer Y, De Wachter R. TREECON for Windows: a software package for the construction and drawing of evolutionary trees for the Microsoft Windows environment. Comput Appl Biosci 1994;; 10: 569–570.
    [Google Scholar]
  33. Wilson MJ, Hall V, Brazier J, Lewis MAO. Evaluation of a phenotypic scheme for the identification of ‘butyrate-producing’ Peptostreptococcus species. J Med Microbiol 2000;; 49: 747–751.
    [Google Scholar]
  34. Kostman JR, Edlind TD, Lipuma JJ, Stull TL. Molecular epidemiology of Pseudomonas cepacia determined by polymerase chain reaction ribotyping. J Clin Microbiol 1992;; 30: 2084–2087.
    [Google Scholar]
  35. Whiley RA, Duke B, Hardie JM, Hall LMC. Heterogeneity among 16S–23S ribosomal RNA intergenic spacers of species within the ‘Streptococcus milleri’ group. Microbiology 1995;; 141: 1461–1467.[CrossRef]
    [Google Scholar]
  36. Murdoch DA, Mitchelmore IJ. The laboratory identification of gram-positive anaerobic cocci. J Med Microbiol 1991;; 34: 295–308.[CrossRef]
    [Google Scholar]
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