Cartridge for performing assays in a closed sample preparation and reaction system

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.

1 . A fluid sample processing cartridge comprising:
 a) a sample preparation module comprising:
 i) a substrate; 
 ii) a sample well formed in said substrate and configured to receive a volume of fluid sample; 
 iii) a closure configured to be selectively placed over said sample well; 
 iv) a first deformable fluid chamber supported on said substrate and configured to hold a fluid therein when in an undeformed state and to collapse upon application of an external compression force to expel at least a portion of the fluid from the first fluid chamber, said first deformable fluid chamber being in fluid communication with said sample well via a channel formed in said substrate; 
 v) a mixing well formed in said substrate, said mixing well being in fluid communication with said sample well via a channel formed in said substrate; 
 vi) a driven mixing apparatus disposed within said mixing well and constructed and arranged to mix the contents of said mixing well; and 
 vii) a first fluid exit port formed in the substrate, said first fluid exit port being in fluid communication with said mixing well via a channel formed in said substrate; and 
   b) a reaction module attached to said sample preparation module and configured to receive a fluid from said sample preparation module via the fluid exit port formed in said sample preparation module, said reaction module comprising:
 i) a top plate comprising
 1) a top surface; 
 2) a raised wall at least partially circumscribing said top surface and in fluid sealing contact with a surface of said sample preparation module to form an interstitial space between the top surface and the surface of said sample preparation module; 
 3) a sample chamber fluidly coupled to the first fluid exit port of the sample preparation module; 
 4) a reagent chamber; and 
 5) a detection chamber; and 
 
 ii) a fluidic processing panel coupled to a bottom surface of said top plate and defining a reaction and processing space between said fluidic processing panel and said top plate, wherein said reaction and processing space is open or openable to said sample chamber, said reaction chamber, and said detection chamber. 
   
     
     
         2 . The fluid sample processing cartridge of  claim 1 , wherein said sample chamber of said reaction module includes an inlet port through which fluid sample enters said sample chamber and including a gap between the first fluid exit port of the sample preparation module and the inlet port of said sample chamber, said gap being open to the interstitial space. 
     
     
         3 . The fluid sample processing cartridge of  claim 1 , wherein the first fluid exit port of the sample preparation module comprises an outlet channel formed through a frustoconical nipple. 
     
     
         4 . The fluid sample processing cartridge of  claim 1 , wherein the reaction module further comprises an electrosensor array disposed in each detection chamber. 
     
     
         5 . The fluid sample processing cartridge of  claim 1 , wherein said top plate of said reaction module further comprises one or more bubble traps, each bubble trap comprising a bubble capture hood open to the reaction and processing space and a vent opening open to the interstitial space. 
     
     
         6 . The fluid sample processing cartridge of  claim 1 , wherein said sample preparation module further comprises:
 a second deformable fluid chamber supported on said substrate and configured to hold a fluid therein when in an undeformed state and to collapse upon application of an external compression force to expel at least a portion the fluid from the fluid chamber; and   a second fluid exit port formed in the substrate, wherein said second fluid exit port is in fluid communication with the second deformable fluid chamber via a channel formed in said substrate, and   wherein said reaction and processing space is fluidly coupled to the second fluid exit port of the sample preparation module.   
     
     
         7 . The fluid sample processing cartridge of  claim 1 , wherein said mixing well comprises:
 a peripheral wall and a floor defining a well; and   a fluid inlet snorkel extending up a side of the peripheral wall extending from the channel communicating the mixing well to the sample well and terminating below a top edge of the peripheral wall.   
     
     
         8 . The fluid sample processing cartridge of  claim 7 , wherein said fluid inlet snorkel extends up an outer surface of the peripheral wall and terminates at an opening formed in the peripheral wall. 
     
     
         9 . The fluid sample processing cartridge of  claim 7 , wherein said mixing well further comprises an exit port comprising one or more openings formed in the floor of the mixing well, wherein said floor tapers downwardly toward said exit port. 
     
     
         10 . The fluid sample processing cartridge of  claim 1 , wherein said driven mixing apparatus comprises a first impeller rotatably disposed within said mixing well and a gear configured to be drivingly engaged by a mating gear of an instrument into which the liquid sample processing cartridge is inserted and to rotate the first impeller when engaged by the mating gear. 
     
     
         11 . The fluid sample processing cartridge of  claim 1 , wherein said sample preparation module further comprises:
 a lysis chamber comprising a plurality of lysis beads, said lysis chamber being formed in said substrate and disposed along the channel connecting said mixing well and said sample well whereby fluid flowing from the sample well to said mixing well will flow through said lysis chamber; and   a bead mixer disposed at least partially within said lysis chamber and constructed and arranged to agitate said lysis beads and fluid flowing through said lysis chamber.   
     
     
         12 . The fluid sample processing cartridge of  claim 11 , further comprising:
 a first optical interface comprising an enlarged portion of the channel connecting said lysis chamber to said sample well; and   a second optical interface comprising an enlarged portion of the channel connecting said lysis chamber to said mixing well.   
     
     
         13 . The fluid sample processing cartridge of  claim 12 , wherein said bead mixer comprises:
 a motor mounted within said substrate; and   a second impeller disposed within said lysis chamber and mounted on an output shaft of said motor.   
     
     
         14 . The fluid sample processing cartridge of  claim 11 , wherein said lysis chamber includes a fluid inlet and a fluid outlet and further comprises a mesh filter disposed over each of said fluid inlet and said fluid outlet and configured to retain said lysis beads within said lysis chamber. 
     
     
         15 . The fluid sample processing cartridge of  claim 1 , wherein said sample preparation module further comprises:
 a pressure port formed in said substrate and configured to couple said substrate to an external fluid pressure source; and   a channel formed in said substrate connecting said pressure port to said sample well.   
     
     
         16 . The fluid sample processing cartridge of  claim 1 , wherein said sample preparation module further comprises:
 a waste chamber formed in said substrate, said waste chamber being in fluid communication with said mixing well via a channel formed in said substrate;   a first externally actuatable control valve disposed within said substrate and constructed and arranged to selectively permit or prevent fluid flow from said mixing well to said waste chamber; and   a second externally actuatable control valve disposed within said substrate and constructed and arranged to selectively permit or prevent fluid flow from said mixing well to said exit port.   
     
     
         17 . The fluid sample processing cartridge of  claim 16 , wherein said sample preparation module further comprises a capture chamber disposed along a channel connecting said mixing well and said waste chamber. 
     
     
         18 . The fluid sample processing cartridge of  claim 1 , wherein said sample preparation module further comprises:
 a passive valve assembly disposed within said substrate and constructed and arranged to be closed and prevent fluid flow from said mixing well when pressure within said mixing well is not higher than a threshold pressure and to open and permit fluid flow from said mixing well when pressure within said mixing well rises above the threshold pressure; and   a pressure port formed in said substrate and in pressure communication with said passive valve assembly by a pressure conduit formed in said substrate, wherein when said pressure port is closed, pressure within said mixing well is allowed to reach the threshold pressure that will open said passive valve assembly and permit fluid flow from said mixing well, and when said pressure port is open pressure within said mixing well cannot reach the threshold pressure so the passive valve assembly is closed.   
     
     
         19 . The fluid sample processing cartridge of  claim 1 , wherein said sample preparation module further comprises a lance blister associated with said deformable fluid chamber; said lance blister being connected or connectable to the associated deformable fluid chamber and containing a bead retained within the lance blister by a breakable septum, wherein said lance blister is configured to collapse upon application of an external compression force to thereby push the bead through the breakable septum. 
     
     
         20 . The fluid sample processing cartridge of  claim 1 , further comprising an external shroud externally enclosing at least a portion of said cartridge. 
     
     
         21 . The fluid sample processing cartridge of  claim 1 , wherein said sample preparation module further comprises a plurality of deformable fluid chambers, each of said fluid chambers containing a substance selected from the group consisting of a lysis buffer, a wash buffer, an oil, a rehydration buffer, target capture beads, and a binding buffer.