Identification of Streptococcus sanguinis with a PCR-generated species-specific DNA probe

Yihong Li, Yaping Pan, Fengxia Qi, Page W. Caufield

Research output: Contribution to journalArticle

Abstract

The objective of the present study was to design a PCR-generated DNA probe and determine the specificity of the probe for the identification of clinical isolates of Streptococcus sanguinis. To do this, we examined over 200 arbitrarily primed PCR (AP-PCR) amplicon patterns obtained with DNA from clinical isolates of S. sanguinis. A 1.6-kb DNA amplicon that was common to all AP-PCR profiles was extracted from agarose gels and then cloned and sequenced. A search for a similar sequence in the GenBank database with the BLASTN program revealed that the 1.6-kb DNA fragment comprised an intergenic region between two housekeeping genes, uncC (proton-translocating ATPase) and murA (UDP-N-acetylglucosamine enolpyruvyl transferase). Three digoxigenin-labeled DNA probes were synthesized on the basis of the sequence of the 1.6-kb fragment: the sequence of probe SSA-1 contained the proton-translocating ATPase (uncC) and the entire intergenic region, the sequence of probe SSA-2 contained only the intergenic region, and the sequence of probe SSA-3 contained an internal region of the murA gene. Dot blot hybridization showed that the three probes displayed signals for hybridization to both S. sanguinis strain ATCC 10556 and the S. sanguinis clinical isolates. Probe SSA-1, however, hybridized to DNA from S. oralis and S. mitis. Probe SSA-3 hybridized to DNA from S. gordonii, S. mitis, S. oralis, S. parasanguinis, and S. vestibularis. The probe SSA-2-specific intergenic region appeared to be specific for S. sanguinis. The results from this study suggest that probe SSA-2 may serve as a species-specific DNA probe for the identification of clinical isolates of S. sanguinis.

Original languageEnglish (US)
Pages (from-to)3481-3486
Number of pages6
JournalJournal of Clinical Microbiology
Volume41
Issue number8
DOIs
StatePublished - Aug 1 2003

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Intergenic DNA
DNA Probes
Streptococcus
Polymerase Chain Reaction
DNA
Proton-Translocating ATPases
Digoxigenin
Essential Genes
Nucleic Acid Databases
Sepharose
Gels
Databases
Genes

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

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Identification of Streptococcus sanguinis with a PCR-generated species-specific DNA probe. / Li, Yihong; Pan, Yaping; Qi, Fengxia; Caufield, Page W.

In: Journal of Clinical Microbiology, Vol. 41, No. 8, 01.08.2003, p. 3481-3486.

Research output: Contribution to journalArticle

Li, Yihong ; Pan, Yaping ; Qi, Fengxia ; Caufield, Page W. / Identification of Streptococcus sanguinis with a PCR-generated species-specific DNA probe. In: Journal of Clinical Microbiology. 2003 ; Vol. 41, No. 8. pp. 3481-3486.
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