Integrated multiplex target analysis

Abstract

This invention provides biochip cartridges and instrument devices for the detection and/or analysis of target analytes from patient samples.

1 - 25 . (canceled) 
     
     
         26 . A biochip cartridge for detecting a target analyte, the biochip cartridge comprising:
 a bottom substrate comprising a printed circuit board, the printed circuit board comprising an amplification zone, the amplification zone comprising (1) at least one lane; and (2) three thermal zones, the first thermal zone being a denature thermal zone and each of the second and third thermal zones being an anneal/extend thermal zone forming two PCR reaction zones per lane, each lane having first and second droplets, the first droplet being disposed in the denature thermal zone and the second droplet being disposed in one of the anneal/extend thermal zones.   
     
     
         27 . A biochip cartridge for detecting a target analyte, the biochip cartridge comprising:
 a bottom substrate comprising a printed circuit board, the printed circuit board comprising an amplification zone, the amplification zone comprising (1) one or more lanes formed by electrowetting pads and (2) a first droplet and a second droplet per lane, wherein at a first time point the first droplet is in a first denature temperature zone and the second droplet is in a first anneal/extend temperature zone, and, at a second time point, the first droplet is in a second anneal/extend temperature zone and the second droplet is in the first denature zone.   
     
     
         28 . A biochip cartridge for detecting a target analyte, the biochip cartridge comprising:
 a bottom substrate comprising a printed circuit board, the printed circuit board comprising an amplification zone, the amplification zone comprising a plurality of lanes, each lane comprising a first and second Polymerase Chain Reaction (PCR) zone.   
     
     
         29 . The biochip cartridge of  claim 28 , wherein the amplification zone comprises one denature zone, a first anneal/extend zone and a second anneal/extend zone. 
     
     
         30 . The biochip cartridge of  claim 28 , wherein the first and second PCR zone each comprise one denature zone and one anneal/extend zone. 
     
     
         31 . The biochip cartridge of  claim 30 , wherein the first and second PCR zone overlap at the denature zone of each PCR zone. 
     
     
         32 . The biochip cartridge of  claim 29 , further comprising first and second droplets in each lane, the first droplet being disposed in the denature zone and the second droplet being disposed in the first anneal/extend zone at a first time point. 
     
     
         33 . The biochip cartridge of  claim 32 , the first droplet being disposed in the second anneal/extend zone and the second droplet being disposed in the denature zone at a second time point. 
     
     
         34 . The biochip cartridge of  claim 29 , further comprising first and second droplets in each lane, wherein the first and second droplets are shuttled independently, the first drop being shuttled between (1) the denature zone and the first anneal/extend zone, and (2) the second drop being shuttled between the denature zone and the second anneal/extend zone. 
     
     
         35 . The biochip cartridge of  claim 29 , further comprising first and second droplets in each lane, wherein drops in a first lane are cycled independently of drops in a second lane. 
     
     
         36 . The biochip cartridge of  claim 34 , wherein the first droplet and the second droplet are shuttled by electrowetting manipulation across electrowetting pads, and wherein at least a portion of the electrowetting pads the first droplet and the second droplet pass over are the same. 
     
     
         37 . The biochip cartridge of  claim 32 , wherein the first droplet and the second droplet are amplified at the same time. 
     
     
         38 . The biochip cartridge of  claim 29 , wherein the denature zone is heated to about 95° C. and wherein the first and the second anneal/extend zones are heated to about 60° C. 
     
     
         39 . The biochip cartridge of  claim 32 , wherein the first droplet and the second droplet comprise reagents for amplification of multiple targets in a single PCR experiment. 
     
     
         40 . The biochip cartridge of  claim 28 , further comprising a single entry lane into the amplification zone. 
     
     
         41 . The biochip cartridge of  claim 28 , further comprising a top plate with a vent spanning the distance between the first and second PCR zone. 
     
     
         42 . The biochip cartridge of  claim 28 , further comprising a detection zone for electrochemical detection of an amplicon. 
     
     
         43 . The biochip cartridge of  claim 28 , further comprising a top plate, wherein the printed circuit board further comprises a sample preparation zone, and wherein a first gap between the top plate and the bottom substrate is smaller over the amplification zone than a second gap between the top plate and the bottom substrate over the sample preparation zone. 
     
     
         44 . The biochip cartridge of  claim 28 , wherein the biochip cartridge is inside a biochip cartridge bay, the biochip cartridge bay comprising one or more movable magnet assemblies, wherein each magnet assembly is in a first position applying substantially no magnetic force to the biochip cartridge. 
     
     
         45 . The biochip cartridge of  claim 30 , wherein the denature zone and the anneal/extend zones are spaced 4500-9000μ apart.