Specific detection of Staphylococcus aureus by real-time PCR

Wednesday, October 26, 2011 — Poster Session IV
2:00 p.m. – 4:00 p.m.	Natcher Conference Center	FDA/CBER	VIROL/MICRO-17

Authors

• K Nagamine
• GC Hung
• SC Lo

Abstract

Modern molecular methods can provide rapid and sensitive alternatives to conventional microbiological methods. In this study, we assessed Insignia program for its ability to identify DNA signatures for S. aureus and the usefulness for the DNA signatures for primer design and their subsequent performance in PCR assays. Initially, the Insignia program produced 1,180 DNA signatures ranged from 149-479 bp in length. A second step to sort and rank these DNA signatures by their similarity with sequences from closely-related using a second computer program dCAS was employed. DNA signatures that were ranked as most dissimilar to closely-related Staphylococcus species were selected and subjected to NCBI-BlastN search to verify the species-specificity of these DNA signatures. Subsequently, only 6 DNA signatures were selected for primer design and for conventional and real-time PCR testing in the laboratory. Preliminary results showed that 3 out of 6 PCR assays designed based on the DNA signatures could achieve high specificity and sensitivity using both conventional and real-time PCR formats. In conclusion, in combination with dCAS and NCBI BlastN, Insignia program can be useful for DNA signature discovery. Such an approach should also be directly applicable to DNA signature discovery for other pathogen of human health concerns.

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