US 12,390,808 B2
Device and method for detecting nucleic acids in biological samples
Timothy Alcorn, Raleigh, NC (US); Steven Dietl, Raleigh, NY (US); Robert Altavela, Webster, NY (US); Michael Carlotta, Lilburn, GA (US); John C. Detter, Melbourne Beach, FL (US); Todd Haran, Bloomfield, NY (US); Michael Murray, Bloomfield, NY (US); Scott Rosebrough, Avon, NY (US); and Jeffrey Serbicki, Holley, NY (US)
Assigned to DEFINITIVE BIOTECHNOLOGIES LLC, Elkridge, MD (US)
Filed by Definitive Biotechnologies LLC, Elkridge, MD (US)
Filed on Jan. 12, 2022, as Appl. No. 17/647,828.
Claims priority of provisional application 63/243,005, filed on Sep. 10, 2021.
Claims priority of provisional application 63/154,217, filed on Feb. 26, 2021.
Claims priority of provisional application 63/136,435, filed on Jan. 12, 2021.
Prior Publication US 2022/0219169 A1, Jul. 14, 2022
Int. Cl. B01L 3/00 (2006.01); B01F 33/302 (2022.01); B01F 33/3033 (2022.01); B01L 7/00 (2006.01); B01L 9/00 (2006.01); B65G 47/80 (2006.01); B82Y 20/00 (2011.01); B82Y 30/00 (2011.01); B82Y 40/00 (2011.01); C12M 1/34 (2006.01); C12M 3/06 (2006.01); C12N 1/14 (2006.01); C12N 1/20 (2006.01); C12Q 1/02 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6844 (2018.01); C12Q 1/6848 (2018.01); C12Q 1/686 (2018.01); G01N 15/10 (2024.01); G01N 15/14 (2024.01); G01N 15/1433 (2024.01); G01N 21/29 (2006.01); G01N 21/65 (2006.01); G01N 33/543 (2006.01); G01N 33/574 (2006.01); G01N 33/58 (2006.01)
CPC B01L 3/502715 (2013.01) [B01L 3/502723 (2013.01); B01L 3/50273 (2013.01); B01L 3/502738 (2013.01); B01L 7/00 (2013.01); C12Q 1/6806 (2013.01); B01L 2200/04 (2013.01); B01L 2200/0684 (2013.01); B01L 2400/06 (2013.01); C12Q 1/6844 (2013.01)] 36 Claims
OG exemplary drawing
 
28. A method of capturing nucleic acids in a biological sample and amplifying the captured nucleic acids therein in a reaction chamber, the method comprising the following steps:
(i) receiving a biological sample in a device, wherein the device comprises a sample port for receiving therein the biological sample;
a lysis chamber including a lysis agent therein;
a mixing chamber for mixing the biological sample and lysis agent into a sample-lysis mixture;
a wash station including a wash solution therein;
an elution station including an eluent therein;
a solid-state membrane located downstream of the mixing chamber, wash station and elution station, and configured to capture nucleic acids in the biological sample passed across the membrane;
a waste chamber located downstream of the solid-state membrane; and
a reaction chamber located downstream of the solid-state membrane;
wherein the sample port, lysis chamber and mixing chamber are configured to mix the sample and lysis agent to form a sample-lysis mixture, pass the sample-lysis mixture across the solid-state membrane to capture nucleic acids in the biological sample therein, and receive the remainder of the sample-lysis mixture in the waste chamber, the wash station is configured to introduce the wash solution across the solid-state membrane to purify nucleic acids captured therein, and receive the wash solution from the solid-state membrane in the waste chamber, and the elution station is configured to pass the eluent across the solid-state membrane, elute captured nucleic acids from the solid-state membrane, and pass the captured nucleic acids into the reaction chamber for amplifying the captured nucleic acids, and
further comprising a sample conduit in fluid communication between each of the sample port, the lysis chamber and the wash station, and the solid-state membrane, wherein the sample conduit includes the mixing chamber therein, the sample port, lysis chamber and sample conduit are configured to mix the sample and lysis agent to form the sample-lysis mixture, pass the sample-lysis mixture across the solid-state membrane to capture nucleic acids in the biological sample therein, and receive any remainder of the sample-lysis mixture in the waste chamber, the wash station is configured to introduce the wash solution into the sample conduit following the sample-lysis mixture, pass the wash solution across the solid-state membrane to purify nucleic acids captured therein, and receive the wash solution from the solid-state membrane in the waste chamber, and the elution station is configured to pass the eluent across the solid-state membrane, elute captured nucleic acids from the solid-state membrane, and pass the captured nucleic acids into the reaction chamber for amplifying the captured nucleic acids, and
(a) a lysis leg extending in fluid communication between the lysis chamber and the sample conduit and configured to direct a flow of the lysis agent from the lysis chamber into the sample conduit, and (b) a wash leg extending in fluid communication between the wash station and the sample conduit at a point upstream relative to the lysis leg and configured to direct a flow of the wash solution from the wash station into the sample conduit behind the sample-lysis mixture;
(ii) passing the biological sample and sample-lysis mixture across solid-state membrane and capturing nucleic acids in the biological sample in the solid-state membrane;
(iii) preventing the flow of the sample-lysis mixture that passes across the solid-state membrane into the reaction chamber, and receiving the remainder of the sample-lysis mixture that passes across the solid-state membrane in the waste chamber;
(iv) passing the wash solution across the solid-state membrane and purifying nucleic acids captured therein;
(v) preventing the flow of the wash solution that passes across the solid-state membrane into the reaction chamber, and receiving the remainder of wash solution that passes across the solid-state membrane in the waste chamber; and
(vi) passing the eluent across the solid-state membrane and eluting captured nucleic acids from the solid-state membrane, directing the eluted captured nucleic acids from the solid-state membrane into the reaction chamber and not into the waste chamber, and amplifying captured nucleic acids therein in the reaction chamber.