Development of species-specific primers for detection ofStreptococcus pyogenesfrom throat swabs

Abstract

A species-specific molecular marker has been developed to detect the human pathogen Streptococcus pyogenes from throat swabs. Streptococcus pyogenes is an important pathogen among Gram-positive organisms. A rapid and simple diagnostic tool is of utmost importance for the identification of this pathogen. The random amplified polymorphic DNA (RAPD) technique was used to differentiate the S. pyogenes strains. A differentially amplified fragment obtained from RAPD profiles was sequenced and characterized, which was developed into a sequence characterized amplified region (SCAR) marker to evaluate the specificity of S. pyogenes from other species of Streptococcus. The sensitivity of the SCAR primers was studied by qualitative PCR and the detection limit was found to be 10(-1) ng of genomic DNA or one to two cells of S. pyogenes. The specificity of the primers was assayed in 270 clinical throat swabs wherein 23 samples turned to be positive, which was highly significant over culture-based methods. This species-specific primer enables accurate detection of S. pyogenes from clinical samples and will be a useful tool in epidemiological studies.