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Anal Chem. 2011 Aug 15;83(16):6349-54. doi: 10.1021/ac2012575. Epub 2011 Jul 14.

Molecular detection of bacterial pathogens using microparticle enhanced double-stranded DNA probes.

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  • 1Department of Aerospace and Mechanical Engineering, University of Arizona, Tucson, Arizona 85721, United States.

Abstract

Rapid, specific, and sensitive detection of bacterial pathogens is essential toward clinical management of infectious diseases. Traditional approaches for pathogen detection, however, often require time-intensive bacterial culture and amplification procedures. Herein, a microparticle enhanced double-stranded DNA probe is demonstrated for rapid species-specific detection of bacterial 16S rRNA. In this molecular assay, the binding of the target sequence to the fluorophore conjugated probe thermodynamically displaces the quencher probe and allows the fluorophore to fluoresce. By incorporation of streptavidin-coated microparticles to localize the biotinylated probes, the sensitivity of the assay can be improved by 3 orders of magnitude. The limit of detection of the assay is as few as eight bacteria without target amplification and is highly specific against other common pathogens. Its applicability toward clinical diagnostics is demonstrated by directly identifying bacterial pathogens in urine samples from patients with urinary tract infections.

PMID:
21718053
[PubMed - indexed for MEDLINE]
PMCID:
PMC4104485
Free PMC Article
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