System and self-metering cartridges for point of care bioassays

Abstract

The invention is directed to devices and methods for performing rapid low-cost bioassays in self-contained disposable cartridges that provide efficient mixing of sample and reactants under a layer of liquid wax. Some embodiments additionally use gravity assisted distribution of sample and assay reagents in conjunction with an appliance containing all necessary valves, pneumatic sources, heat sources and detection stations.

1 - 20 . (canceled) 
     
     
         21 . A bioassay cartridge, comprising:
 a sample chamber capable of receiving a biological sample comprising at least one polynucleotide;   a lysis reservoir in fluid communication with the sample chamber;   a detection chamber in fluid communication with the sample chamber; and
 at least one bioassay reagent, wherein the at least one bioassay reagent is capable of replicating the at least one polynucleotide via isothermal polynucleotide amplification. 
   
     
     
         22 . The bioassay cartridge of  claim 21 , wherein the lysis reservoir contains a lysis buffer. 
     
     
         23 . The bioassay cartridge of  claim 22 , wherein the lysis buffer is capable of disrupting viral protein coatings of viral particles. 
     
     
         24 . The bioassay cartridge of  claim 22 , wherein the lysis buffer comprises at least one chaotropic agent. 
     
     
         25 . The bioassay cartridge of  claim 22 , wherein the lysis buffer comprises at least one nuclease inhibitor. 
     
     
         26 . The bioassay cartridge of  claim 21 , wherein the at least one bioassay reagent comprises at least one set of primers for amplifying at least one internal standard polynucleotide. 
     
     
         27 . The bioassay cartridge of  claim 21 , wherein the at least one bioassay reagent comprises at least two primers capable of forming a duplex with the at least one polynucleotide. 
     
     
         28 . The bioassay cartridge of  claim 21 , further comprising a detection chamber vent port in fluid communication with the detection chamber. 
     
     
         29 . The bioassay cartridge of  claim 28 , wherein the detection chamber vent port is capable of being sealingly connected to a vacuum source. 
     
     
         30 . The bioassay cartridge of  claim 29 , wherein applying a vacuum to the detection chamber vent port permits movement of the at least one polynucleotide into the detection chamber. 
     
     
         31 . The bioassay cartridge of  claim 21 , wherein at least some of the at least one bioassay reagent is contained in the detection chamber. 
     
     
         32 . The bioassay cartridge of  claim 30 , wherein the at least one bioassay reagent contained in the detection chamber comprises at least two primers. 
     
     
         33 . The bioassay cartridge of  claim 21 , wherein at least some of the at least one bioassay reagent is contained in the sample chamber. 
     
     
         34 . The bioassay cartridge of  claim 32 , wherein the at least one bioassay reagent contained in the sample chamber comprises an RNase inhibitor. 
     
     
         35 . The bioassay cartridge of  claim 21 , further comprising a sample chamber vent port in fluid communication with the sample chamber. 
     
     
         36 . The bioassay cartridge of  claim 35 , wherein the sample chamber vent port is capable of being sealingly connected to a pressure source. 
     
     
         37 . The bioassay cartridge of  claim 36 , wherein applying pressure at the sample chamber vent port permits mixing of the at least one polynucleotide and the lysis buffer in the sample chamber. 
     
     
         38 . The bioassay cartridge of  claim 21 , wherein the resulting amplified at least one polynucleotide is capable of fluorescent optical detection. 
     
     
         39 . The bioassay cartridge of  claim 21 , wherein the isothermal polynucleotide amplification comprises loop-mediated isothermal (LAMP) amplification.