title: Rapid detection of urinary tract infections using isotachophoresis and molecular beacons.
abstract: We present a novel assay for rapid detection and identification of bacterial urinary tract infections using isotachophoresis (ITP) and molecular beacons. We applied on-chip ITP to extract and focus 16S rRNA directly from bacterial lysate and used molecular beacons to achieve detection of bacteria specific sequences. We demonstrated detection of E. coli in bacteria cultures as well as in patient urine samples in the clinically relevant range 1E6-1E8 cfu/mL. For bacterial cultures we further demonstrate quantification in this range. The assay requires minimal sample preparation (a single centrifugation and dilution), and can be completed, from beginning of lysing to detection, in under 15 min. We believe that the principles presented here can be used for design of other rapid diagnostics or detection methods for pathogenic diseases.
authors.affiliation: Department of Mechanical Engineering, Stanford University, California 94305, USA.
authors.forename: M
authors.initials: M
authors.lastname: Bercovici
authors.forename: G V
authors.initials: GV
authors.lastname: Kaigala
authors.forename: K E
authors.initials: KE
authors.lastname: Mach
authors.forename: C M
authors.initials: CM
authors.lastname: Han
authors.forename: J C
authors.initials: JC
authors.lastname: Liao
authors.forename: J G
authors.initials: JG
authors.lastname: Santiago
mesh_terms: D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D057625:Isotachophoresis / Q000379:methods*; D046210:Microfluidic Analytical Techniques / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D014552:Urinary Tract Infections / Q000175:diagnosis* / Q000382:microbiology
chemical_list: D005456:Fluorescent Dyes; D012336:RNA, Ribosomal, 16S