title: Molecular detection of bacterial pathogens using microparticle enhanced double-stranded DNA probes.
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.
authors: Department of Aerospace and Mechanical Engineering, University of Arizona, Tucson, Arizona 85721, United States.
authors: Reza
authors: R
authors: Riahi
authors: Kathleen E
authors: KE
authors: Mach
authors: Ruchika
authors: R
authors: Mohan
authors: Joseph C
authors: JC
authors: Liao
authors: Pak Kin
authors: PK
authors: Wong
mesh_terms: D001681:Biological Assay; D001710:Biotin / Q000378:metabolism; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D015342:DNA Probes / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000254:growth & development; D005456:Fluorescent Dyes / Q000032:analysis; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D011549:Pseudomonas / Q000737:chemistry* / Q000254:growth & development; D012336:RNA, Ribosomal, 16S / Q000032:analysis*; D012680:Sensitivity and Specificity; D057790:Staphylococcus saprophyticus / Q000737:chemistry* / Q000254:growth & development; D019809:Streptavidin / Q000378:metabolism; D014552:Urinary Tract Infections / Q000175:diagnosis* / Q000235:genetics / Q000382:microbiology
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D012336:RNA, Ribosomal, 16S; D001710:Biotin; D004247:DNA; D019809:Streptavidin