Miniaturized Lateral Flow Device for Rapid and Sensitive Detection of Proteins or Nucleic Acids

摘要

The invention provides miniaturized lateral flow chromatographic and lateral flow chromatographic microarray devices (LFM). The miniaturization of lateral flow nucleic acid detection achieved by the present invention offers reduced reagent use, femtomole sensitivity, excellent linear dynamic range, and rapid detection. Moreover, the small feature sizes of capture oligonucleotides renders the potential information capacity of the platform comparable to more traditional spotted fluorescence microarrays as well as improving sensitivity. The LFM devices exemplified herein enable analytes to be detected within 10 seconds from the time of sample introduction to the LFM device. Sample volumes may be as low as about 10 microliters, significantly reducing assay costs and ameliorating reagent storage logistics. Additionally, the miniaturization of lateral flow opens the door to highly multiplexed assays, allowing many proteins or nucleic acids to be detected in a single assay.

主权项

1. A method for quantitatively detecting the presence of one or more target nucleic acids in a fluid sample, the method comprising:
immobilizing first capture oligonucleotides on a first region of a microporous membrane; immobilizing second capture oligonucleotides on a second region of a microporous membrane; hybridizing labeled colorimetric detection oligonucleotides to a complementary first sequence of the target nucleic acid; directly hybridizing the first capture oligonucleotides to a complementary second sequence of a first group of the target nucleic acids; directly hybridizing the second capture oligonucleotides to a complementary third sequence of a second group of the target nucleic acids, wherein the first capture oligonucleotides and the second capture oligonucleotides each display signal linearity over a different concentration range of the target nucleic acid; measuring an intensity of a first optical or colorimetric signal produced on the first region; measuring an intensity of a second optical or colorimetric signal produced on the second region; determining if the intensity of the first signal and/or the second signal is in the linear range of the first capture oligonucleotide and second capture oligonucleotide respectively; and calculating the concentration of the target nucleic acid in the fluid sample using the intensity of the first signal and/or the second signal if the intensity is in the respective linear range.