A capture oligonucleotide-based surface plasmon resonance assay was developed for the specific detection of tmRNA from the gram-positive pathogen Streptococcus pneumoniae. Candidate species-specific probes were initially screened on microarrays using labeled in vitro transcribed tmRNA beforebeing subjected to further analysis on a Biacore 2000 biosensor using a variety of unlabeled tmRNA-containing samples. Previous work had demonstrated that there is approximately 1,000 copies/cfu of tmRNA in S. pneumoniae. The natural amplification of this target has the potential to improve the detection capability of a nucleic acid diagnostics assay. In this study, S. pneumoniae-specific probes were designed that enabled the detection of 10 exp12 cRNA copies of the tmRNA target on the Biacore instrument. When evaluated in combination with nucleic acid sequence-based amplification, specific detection of tmRNA from a single S. pneumoniae cell was demonstrated.
|Number of pages||12|
|Journal||Journal of Rapid Methods & Automation in Microbiology|
|Publication status||Published - Sep 2008|