Patent ID: 7604938
Filing Date: 2009-10-20
Classification: B01L,C12Q,G01N

Abstract:
1. A method of detecting the absence or presence of one or more organisms in one or more samples, comprising the steps of: (a) providing a microfluidic device comprising a microfluidic reagent assembly area which forms two or more sample plugs, said microfluidic reagent assembly area being in fluid communication with at least one microfluidic inline channel that runs through an amplification area and a detection area, said microfluidic reagent assembly area comprising: wherein said microfluidic reagent assembly area is controlled to form sample plugs that comprise the same or different genomic material and the same or different primers; (b) acquiring samples that may contain an organism; (c) selecting one or more samples to input into said sample droplet forming area of the device; (d) isolating at least one copy of the genomic material if present from each of the selected samples in the sample droplet forming area; (e) introducing at least one sample droplet of each of the selected samples into the microfluidic inline channel through the sample droplet microfluidic path; (f) driving the sample droplets along the microfluidic inline channel through the reagent assembly area; (g) selecting two or more primer plugs to input into the primer plug microfluidic path of the device; (h) assembling the selected primer plugs in the primer plug assembly area, wherein each primer plug comprises amplification reagents and at least one primer, and wherein the primer plugs contain the same primers or different primers; (i) introducing the primer plugs into the microfluidic inline channel through the primer plug microfluidic path; (j) forming two or more sample plugs in the microfluidic inline channel by mixing the sample droplets with the primer plugs in the microfluidic inline channel in the sample plug mixing area, wherein the sample plugs comprise the same or different genomic material and the same or different primers; (k) driving the sample plugs along the microfluidic inline channel through the amplification area; (l) amplifying the genomic material, if present, in each of the sample plugs in the microfluidic inline channel in the amplification area of the device; (m) driving the sample plugs along the microfluidic inline channel through the detection area; and (n) detecting the absence or presence of amplified products in each of the sample plugs, wherein the presence of amplified products for an organism is indicative of the presence of the organism in the sample.