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. 1996 Jul;34(7):1717–1721. doi: 10.1128/jcm.34.7.1717-1721.1996

Rapid species identification of "Streptococcus milleri" strains by line blot hybridization: identification of a distinct 16S rRNA population closely related to Streptococcus constellatus.

J A Jacobs 1, C S Schot 1, A E Bunschoten 1, L M Schouls 1
PMCID: PMC229101  PMID: 8784576

Abstract

A collection of 399 "Streptococcus milleri" strains were identified to the species level by the use of a line blot assay. Their PCR-amplified partial 16S rRNA gene sequences were hybridized with species-specific 5'-biotinylated oligonucleotide probes homologous to the bp 213 to 231 regions of the 16S rRNA gene sequences of the type strains Streptococcus anginosus ATCC 33397, Streptococcus constellatus ATCC 27823, and Streptococcus intermedius ATCC 27335. The hybridization results were compared with the reference phenotypic identification method data (R. A. Whiley, H. Fraser, J. M. Hardie, and D. Beighton, J. Clin. Microbiol. 28:1497-1501, 1990). Most strains (357 of 399 [89.5%]) reacted unambiguously with only one probe. However, 42 of the 399 strains (10.5%) reacted with both the S. constellatus- and S. intermedius-specific probes; 41 of them were phenotypically identified as S. constellatus. These dually reactive strains hybridized with a 5'-biotinylated probe based on the bp 213 to 231 region of the 16S rRNA gene sequence of one of two species. Analysis of the 5' ends of the 16S rRNA gene sequences (487 bp) demonstrated that the dually reactive strains represent a distinct rRNA population sharing 98.1% sequence similarity with S. constellatus. Phenotypic consistency between the dually reactive strains and the S. constellatus strains was not demonstrated. Line blot hybridization proved to be a simple and inexpensive method to screen large numbers of strains for genetic relatedness, and it allowed the detection of a distinct 16S rRNA type within the "S. milleri" group.

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Selected References

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  1. Bentley R. W., Leigh J. A., Collins M. D. Intrageneric structure of Streptococcus based on comparative analysis of small-subunit rRNA sequences. Int J Syst Bacteriol. 1991 Oct;41(4):487–494. doi: 10.1099/00207713-41-4-487. [DOI] [PubMed] [Google Scholar]
  2. Bergman S., Selig M., Collins M. D., Farrow J. A., Baron E. J., Dickersin G. R., Ruoff K. L. "Streptococcus milleri" strains displaying a gliding type of motility. Int J Syst Bacteriol. 1995 Apr;45(2):235–239. doi: 10.1099/00207713-45-2-235. [DOI] [PubMed] [Google Scholar]
  3. Bergmans A. M., Groothedde J. W., Schellekens J. F., van Embden J. D., Ossewaarde J. M., Schouls L. M. Etiology of cat scratch disease: comparison of polymerase chain reaction detection of Bartonella (formerly Rochalimaea) and Afipia felis DNA with serology and skin tests. J Infect Dis. 1995 Apr;171(4):916–923. doi: 10.1093/infdis/171.4.916. [DOI] [PubMed] [Google Scholar]
  4. Gossling J. Occurrence and pathogenicity of the Streptococcus milleri group. Rev Infect Dis. 1988 Mar-Apr;10(2):257–285. doi: 10.1093/clinids/10.2.257. [DOI] [PubMed] [Google Scholar]
  5. Jacobs J. A., Pietersen H. G., Stobberingh E. E., Soeters P. B. Streptococcus anginosus, Streptococcus constellatus and Streptococcus intermedius. Clinical relevance, hemolytic and serologic characteristics. Am J Clin Pathol. 1995 Nov;104(5):547–553. doi: 10.1093/ajcp/104.5.547. [DOI] [PubMed] [Google Scholar]
  6. Jacobs J. A., Schouten H. C., Stobberingh E. E., Soeters P. B. Viridans streptococci isolated from the bloodstream. Relevance of species identification. Diagn Microbiol Infect Dis. 1995 Jul;22(3):267–273. doi: 10.1016/0732-8893(95)00137-y. [DOI] [PubMed] [Google Scholar]
  7. Kaufhold A., Podbielski A., Baumgarten G., Blokpoel M., Top J., Schouls L. Rapid typing of group A streptococci by the use of DNA amplification and non-radioactive allele-specific oligonucleotide probes. FEMS Microbiol Lett. 1994 Jun 1;119(1-2):19–25. doi: 10.1111/j.1574-6968.1994.tb06861.x. [DOI] [PubMed] [Google Scholar]
  8. Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Piscitelli S. C., Shwed J., Schreckenberger P., Danziger L. H. Streptococcus milleri group: renewed interest in an elusive pathogen. Eur J Clin Microbiol Infect Dis. 1992 Jun;11(6):491–498. doi: 10.1007/BF01960802. [DOI] [PubMed] [Google Scholar]
  10. Rijpkema S. G., Molkenboer M. J., Schouls L. M., Jongejan F., Schellekens J. F. Simultaneous detection and genotyping of three genomic groups of Borrelia burgdorferi sensu lato in Dutch Ixodes ricinus ticks by characterization of the amplified intergenic spacer region between 5S and 23S rRNA genes. J Clin Microbiol. 1995 Dec;33(12):3091–3095. doi: 10.1128/jcm.33.12.3091-3095.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ruoff K. L., Kunz L. J., Ferraro M. J. Occurrence of Streptococcus milleri among beta-hemolytic streptococci isolated from clinical specimens. J Clin Microbiol. 1985 Aug;22(2):149–151. doi: 10.1128/jcm.22.2.149-151.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ruoff K. L. Streptococcus anginosus ("Streptococcus milleri"): the unrecognized pathogen. Clin Microbiol Rev. 1988 Jan;1(1):102–108. doi: 10.1128/cmr.1.1.102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Weisburg W. G., Barns S. M., Pelletier D. A., Lane D. J. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol. 1991 Jan;173(2):697–703. doi: 10.1128/jb.173.2.697-703.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Whiley R. A., Beighton D. Emended descriptions and recognition of Streptococcus constellatus, Streptococcus intermedius, and Streptococcus anginosus as distinct species. Int J Syst Bacteriol. 1991 Jan;41(1):1–5. doi: 10.1099/00207713-41-1-1. [DOI] [PubMed] [Google Scholar]
  15. Whiley R. A., Fraser H., Hardie J. M., Beighton D. Phenotypic differentiation of Streptococcus intermedius, Streptococcus constellatus, and Streptococcus anginosus strains within the "Streptococcus milleri group". J Clin Microbiol. 1990 Jul;28(7):1497–1501. doi: 10.1128/jcm.28.7.1497-1501.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Woese C. R. Bacterial evolution. Microbiol Rev. 1987 Jun;51(2):221–271. doi: 10.1128/mr.51.2.221-271.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]

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