Processing Cartridge and Method for Detecting a Pathogen in a Sample

Abstract

In one embodiment, a multiplex fluid processing cartridge includes a sample well, a deformable fluid chamber, a mixing well with a mixer disposed therein, a lysis chamber including a lysis mixer, an electrowetting grid for microdroplet manipulation, and electrosensor arrays configured to detect analytes of interest. An instrument for processing the cartridge is configured to receive the cartridge and to selectively apply thermal energy, magnetic force, and electrical connections to one or more discrete locations on the cartridge and is further configured to compress the deformable chamber(s) in a specified sequence.

What is claimed is: 
     
         1 . A method of motivating a sample through a cartridge, the method comprising:
 (a) motivating the sample on a first substrate using microfluidics; and   (b) motivating the sample on a second substrate using electrowetting, thereby motivating a sample through a cartridge, wherein (a) is not accomplished by electrowetting.   
     
     
         2 . The method of  claim 1 , further comprising, after (b), motivating the sample into an electrosensor array located on the second substrate. 
     
     
         3 . The method of  claim 1 , wherein (a) comprises pumping the sample from a first location to a second location. 
     
     
         4 . The method of  claim 1 , wherein (a) comprises compressing a fluid-filled deformable compartment supported on the first substrate, thereby generating pressure to move the sample. 
     
     
         5 . The method of  claim 1 , wherein (a) comprises introducing magnetic beads to the sample and applying a magnetic force to move the sample. 
     
     
         6 . The method of  claim 1 , wherein (a) comprises introducing negatively-charged capture beads to the first substrate to facilitate adsorption of positively-charged nucleic acids from the sample to a surface of the negatively-charged capture beads, thereby moving the sample. 
     
     
         7 . The method of  claim 1 , further comprising introducing process fluids into the first substrate by compressing a fluid-filled deformable compartment supported on the first substrate. 
     
     
         8 . The method of  claim 1 , further comprising:
 (c) retaining on the second substrate by electrowetting manipulation an aliquot of the sample comprising: magnetic beads with DNA material bound thereto and wash solution; and   (d) pulling the magnetic beads out of the wash solution using magnetic forces.   
     
     
         9 . The method of  claim 1 , wherein (a) comprises motivating the sample by pumping the sample, generating pressure in a microfluidic channel, applying magnetic forces to the sample, absorbing the sample, applying rotary forces to the sample, or combinations thereof. 
     
     
         10 . The method of  claim 1 , wherein (a) comprises motivating the sample by pumping the sample, generating pressure in a microfluidic channel, applying magnetic forces to the sample, absorbing the sample, and applying rotary forces to the sample. 
     
     
         11 . The method of  claim 1 , further comprising, before (b), passing the sample through an interstitial space between the first substrate and the second substrate, wherein the interstitial space forms a void. 
     
     
         12 . A method of motivating a sample through a cartridge, the method comprising:
 (a) motivating the sample on a first substrate using a first microfluidic technique; and   (b) motivating the sample on a second substrate using electrowetting, wherein the first microfluidic technique and electrowetting are different, thereby motivating a sample through a cartridge.   
     
     
         13 . The method of  claim 12 , wherein (a) comprises motivating the sample by pumping the sample, generating pressure in a microfluidic channel, applying magnetic forces to the sample, absorbing the sample, applying rotary forces to the sample, or combinations thereof. 
     
     
         14 . The method of  claim 12 , further comprising receiving a signal from an optical detector indicating an end of fluid flow through an optical sensing chamber and in response to the signal deactivating a rotary mixer motor. 
     
     
         15 . The method of  claim 12 , further comprising, after (b), on the second substrate, retaining magnetic target capture beads and target analyte bound thereto by magnetic forces while flowing a remainder of contents of the sample into a waste chamber by electrowetting manipulation. 
     
     
         16 . A method of motivating a sample through a cartridge, the method comprising:
 (a) motivating the sample on a first substrate using microfluidics;   (b) motivating the sample out of the first substrate into a sample outlet;   (c) motivating the sample out of the sample outlet and into a sample inlet, thereby allowing gas bubbles to escape into an interstitial space;   (d) motivating the sample out of the sample inlet and onto the second substrate; and   (e) motivating the sample through the second substrate using electrowetting, thereby motivating a sample through a cartridge.   
     
     
         17 . The method of  claim 16 , wherein (a) comprises motivating the sample by pumping the sample, generating pressure in a microfluidic channel, applying magnetic forces to the sample, absorbing the sample, applying rotary forces to the sample, or combinations thereof. 
     
     
         18 . The method of  claim 16 , further comprising, after (e), on the second substrate, motivating the sample under a bubble trap allowing bubbles to escape into the interstitial space between the first substrate and the second substrate. 
     
     
         19 . The method of  claim 16 , further comprising applying a voltage to detection electrodes in an electrosensor array on the second substrate. 
     
     
         20 . The method of  claim 16 , further comprising introducing process fluids into the second substrate by motivating fluid through an inlet channel on the first substrate, out an outlet port on the first substrate, and in an inlet port on the second substrate. 
     
     
         21 . A method for detecting a pathogen in a sample, the method comprising:
 loading a sample into a cartridge, the cartridge comprising a first substrate and a second substrate;   extracting nucleic acids from the sample in the first substrate;   transferring the sample to the second substrate; and   detecting a pathogen, if present, in the sample in the second substrate.