US 11,872,554 B2
Microfluidic devices and related methods
Alexander Schenk zu Schweinsberg, Jena (DE); Austin Matthew Derfus, Solana Beach, CA (US); Justin Davidson, San Diego, CA (US); Karthikeyan Kumaravadivelu, San Diego, CA (US); Maulik Vinod Patel, La Crescenta, CA (US); Olaf Piepenburg, Saffron Walden (GB); Catherine Jean Greenwood, Sawbridgeworth (GB); and Oliver Nentwich, Cambridge (GB)
Assigned to ABBOTT DIAGNOSTICS SCARBOROUGH, INC., Scarborough, ME (US)
Filed by ABBOTT DIAGNOSTICS SCARBOROUGH, INC., Scarborough, ME (US)
Filed on Mar. 9, 2020, as Appl. No. 16/812,961.
Application 16/812,961 is a division of application No. 15/906,899, filed on Feb. 27, 2018, granted, now 10,632,464.
Claims priority of provisional application 62/464,576, filed on Feb. 28, 2017.
Prior Publication US 2020/0391209 A1, Dec. 17, 2020
Int. Cl. G01N 1/00 (2006.01); B01L 3/00 (2006.01); G01N 1/14 (2006.01); G01N 35/00 (2006.01); G01N 1/02 (2006.01); G01N 1/38 (2006.01); G01N 35/10 (2006.01)
CPC B01L 3/502715 (2013.01) [B01L 3/5029 (2013.01); G01N 1/14 (2013.01); G01N 35/00069 (2013.01); B01L 2200/027 (2013.01); B01L 2200/082 (2013.01); B01L 2300/0816 (2013.01); B01L 2300/0864 (2013.01); B01L 2300/0867 (2013.01); B01L 2400/0478 (2013.01); B01L 2400/0655 (2013.01); G01N 1/38 (2013.01); G01N 2001/028 (2013.01); G01N 2035/00158 (2013.01); G01N 2035/1034 (2013.01)] 18 Claims
OG exemplary drawing
 
1. A method comprising:
providing a microfluidic device comprising: a first reaction chamber comprising a first magnet; and a second reaction chamber comprising a second magnet;
mixing a sample fluid comprising a target nucleic acid with a first set of recombinase polymerase amplification (RPA) reagents;
providing the sample fluid comprising the target nucleic acid and the RPA reagents to flail the microfluidic device, the target nucleic acid comprising a target polynucleotide sequence; and
amplifying the target polynucleotide sequence under isothermal conditions, wherein the amplifying comprises:
performing a first round of amplification on the target polynucleotide sequence in all the first reaction chamber to yield a first amplification product comprising a first amplified polynucleotide sequence;
mixing a portion of the first amplification product with a second set of RPA reagents in flail the second reaction chamber; and
performing a second round of amplification on the first amplified polynucleotide sequence in the second reaction chamber to yield a second amplification product comprising a second amplified polynucleotide sequence,
wherein the second amplified polynucleotide sequence comprises a smaller sequence completely contained within the first amplified polynucleotide sequence produced during the first round of amplification,
wherein the second amplification product is detected in less than approximately 30 minutes after providing the sample to the microfluidic device.