Lateral flow microarrays: a novel platform for rapid nucleic acid detection based on miniaturized lateral flow chromatography.
Widely used nucleic acid assays are poorly suited for field deployment where access to laboratory instrumentation is limited or unavailable. The need for field deployable nucleic acid detection demands inexpensive, facile systems without sacrificing information capacity or sensitivity. Here we describe a novel microarray platform capable of rapid, sensitive nucleic acid detection without specialized instrumentation. The approach is based on a miniaturized lateral flow device that makes use of hybridization-mediated target capture. The miniaturization of lateral flow nucleic acid detection provides multiple advantages over traditional lateral flow devices. Ten-microliter sample volumes reduce reagent consumption and yield analyte detection times, excluding sample preparation and amplification, of <120 s while providing sub-femtomole sensitivity. Moreover, the use of microarray technology increases the potential information capacity of lateral flow. Coupled with a hybridization-based detection scheme, the lateral flow microarray (LFM) enables sequence-specific detection, opening the door to highly multiplexed implementations for broad-range assays well suited for point-of-care and other field applications. The LFM system is demonstrated using an isothermal amplification strategy for detection of Bacillus anthracis, the etiologic agent of anthrax. RNA from as few as two B. anthracis cells was detected without thermocycling hardware or fluorescence detection systems.
Carter
Darren J
DJ
Biosciences Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545, USA.
Cary
R Bruce
RB
D000483:Alleles; D001408:Bacillus anthracis / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000235:genetics; D002845:Chromatography / Q000379:methods*; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008904:Miniaturization; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D012329:RNA, Bacterial / Q000032:analysis; D015534:Trans-Activators / Q000235:genetics
D001426:Bacterial Proteins; C099480:PlcR protein, Bacillus; D012329:RNA, Bacterial; D015534:Trans-Activators