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Rapid typing of group A streptococci by the use of DNA amplification and non-radioactive allele-specific oligonucleotide probes

A. Kaufhold, A. Podbielski, G. Baumgarten, M. Blokpoel, J. Top, L. Schouls
DOI: http://dx.doi.org/10.1111/j.1574-6968.1994.tb06861.x 19-25 First published online: 1 June 1994

Abstract

Because of the allelic variations within the M protein gene (emm gene) of group A streptococci, reliable typing of this important human pathogen can be accomplished by the use of emm gene-specific oligonucleotide probes. Two technical modifications (a reverse dot blot and a reverse line blot hybridization assay) of a novel approach for the type-specific identification of emm genes have been developed. Both procedures involved amplification of an emm gene by polymerase chain reaction. The non-radioactively labeled amplicon was subsequently hybridized to a membrane carrying an array of immobilized emm gene-specific oligonucleotide probes, thus allowing the simultaneous analysis of the gene polymorphism in a single hybridization reaction. The feasibility of these rapid and easy to perform methods was shown for the unequivocal identification of reference strains and clinical isolates belonging to 16 different M serotypes.

Key words
  • Streptococcus pyogenes
  • M protein
  • Polymerase chain reaction
  • Hybridization assay
  • Gene polymorphism
  • Epidemiology

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