# US 2014/0194305 A1 · Integrated multiplex target analysis

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- Number: US 2014/0194305 A1
- Document ID: US20140194305A1
- Status: Published application
- Kind: A1
- Application number: 14/062,865
- Filed: 2013-10-24
- Published: 2014-07-10
- Earliest claimed: 2012-10-24
- Assignees: GenMark Diagnostics, Inc.
- Inventors: Jon Faiz KAYYEM; Jay SRINIVASAN
- Web page: https://patentgrep.com/patent/US20140194305A1

## CPC

- C12Q 1/6825: Nucleic acid detection involving sensors
- B01L 3/502715: {characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces}
- B01L 3/502738: {characterised by integrated valves (throttle valves in microfluidic sample containers B01L3/502746)}
- B01L 3/502784: {specially adapted for droplet or plug flow, e.g. digital microfluidics}
- B01L 3/523: {with means for closing or opening}
- B01L 7/52: {with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples}
- B01L 9/527: {for microfluidic devices, e.g. used for lab-on-a-chip}
- B01L 2200/025: Align devices or objects to ensure defined positions relative to each other
- B01L 2200/10: Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
- B01L 2200/16: Reagents, handling or storing thereof
- B01L 2300/021: Identification, e.g. bar codes
- B01L 2300/0672: Integrated piercing tool
- B01L 2300/0816: Cards, e.g. flat sample carriers usually with flow in two horizontal directions
- B01L 2300/0867: Multiple inlets and one sample wells, e.g. mixing, dilution
- B01L 2300/1822: using Peltier elements
- B01L 2300/1827: using resistive heater
- B01L 2400/0427: Electrowetting
- B01L 2400/0481: squeezing of channels or chambers
- B01L 2400/0683: mechanically breaking a wall or membrane within a channel or chamber
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE

## Abstract

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

## Claims

### Claim 1

A biochip cartridge comprising:
a) a bottom substrate comprising a printed circuit board (PCB) comprising:
i) an electrowetting grid of electrodes forming a droplet pathway comprising a plurality of electrowetting pads;
ii) an array of detection electrodes accessible to said droplet pathway, each comprising a self-assembled monolayer and a capture probe;
iii) a plurality of interconnections from said electrowetting grid and said detection electrodes; and
b) a top plate parallel to said bottom substrate and mated thereto to form a reaction chamber.

### Claim 2 (depends on 1)

A biochip cartridge according to claim 1, wherein said top plate comprises a conductive surface.

### Claim 3 (depends on 1)

A biochip cartridge according to claim 1, wherein said top plate comprises at least one fluid hole spatially corresponding to at least one of said electrowetting pads.

### Claim 4 (depends on 1)

A biochip cartridge according to claim 1, wherein at least one of said electrowetting pads comprises a dried assay reagent.

### Claim 5 (depends on 1)

A biochip cartridge according to claim 1, wherein said array of detection electrodes is in fluid communication with said droplet pathway.

### Claim 6 (depends on 1)

A biochip cartridge according to claim 1, wherein said biochip cartridge further comprises a liquid regent module (LRM) comprising:
i) a plurality of blisters comprising fluid assay reagents;
ii) a plurality of passageways in fluid communication with said blisters; and
iii) a sample inlet port in fluid connection with said reaction chamber.

### Claim 7 (depends on 1)

A biochip cartridge according to claim 1, further comprising an external housing comprising a latched cover for sealing said sample inlet port.

### Claim 8 (depends on 7)

A biochip cartridge according to claim 7, wherein said external housing further comprises electronic connections from said interconnections.

### Claim 9 (depends on 1)

A biochip cartridge according to claim 1, wherein said bottom substrate further comprises at least one amplification pathway comprising a plurality of electrowetting pads.

### Claim 10 (depends on 1)

A biochip cartridge according to claim 1, wherein said bottom substrate further comprises a plurality of amplification pathways, each comprising a plurality of electrowetting pads.

### Claim 11 (depends on 6)

A biochip cartridge according to claim 6, wherein said LRM further comprises an aliquot of magnetic capture beads.

### Claim 12 (depends on 6)

A biochip cartridge according to claim 6, wherein at least one of said fluid passageways is configured to dispense a fluid assay reagent at a location remote from a blister comprising said reagent.

### Claim 13 (depends on 7)

A biochip cartridge according to claim 7, wherein said external housing is asymmetrically shaped.

### Claim 14 (depends on 4)

A biochip cartridge according to claim 4, wherein said dried assay reagent is selected from the group consisting of: dNTPs, PCR primers, a nucleic acid polymerase, an exonuclease, labelled signaling probes, and a combination thereof.

### Claim 15 (depends on 7)

A biochip cartridge according to claim 7, wherein said external housing further comprises a cartridge identification tag.

### Claim 16 (depends on 6)

A biochip cartridge according to claim 6, wherein at least one of said fluid assay reagents contained in said plurality of LRM blisters is selected from the group consisting of: water, an immiscible fluid, a lysis buffer, a binding buffer, an elution buffer, and a reconstitution buffer.

### Claim 17

A biochip cartridge comprising:
a) a bottom substrate comprising:
i) a printed circuit board (PCB) comprising:
1) an electrowetting grid of electrodes forming a droplet pathway comprising electrowetting pads, wherein at least one of said pads comprises a dried reagent;
2) an array of detection electrodes, each comprising a self-assembled monolayer and a capture probe, wherein said detection electrodes are accessible to said droplet pathway;
3) a plurality of interconnections from said electrowetting grid and said detection electrodes;
b) a top plate parallel to said bottom substrate and mated thereto to form a reaction chamber, said top plate comprising at least one fluid hole spatially corresponding to said at least one pad comprising said dried reagent;
c) a liquid reagent module (LRM) comprising:
i) a plurality of blisters comprising fluid assay reagents;
ii) a plurality of passageways in fluid connection with said blisters;
iii) a sample inlet port in fluid connection with said reaction chamber; and
d) an external housing comprising:
i) a latched cover for sealing said sample inlet port;
ii) a plurality of electronic connections from said interconnections.

### Claim 18 (depends on 17)

A biochip cartridge according to claim 17, wherein said top plate comprises a conductive surface.

### Claim 19 (depends on 1)

A method of detecting a plurality of target nucleic acids in a sample comprising:
a) adding said sample to a cartridge according to claim 1,
b) executing assay operations on said sample to detect target nucleic acids, wherein said operations comprise:
i) mixing said sample with a lysis buffer;
ii) adding a binding buffer and capture beads to said sample;
iii) mixing said capture beads to facilitate target binding;
iv) washing said capture beads to remove non-target components;
v) eluting said target nucleic acids from said capture beads;
vi) amplifying said target nucleic acids to form a plurality of amplicons;
vii) adding signaling probes to said amplicons to form a plurality of hybridization complexes;
viii) binding said hybridization complexes to said capture probes on said detection electrodes to form a plurality of assay complexes; and
ix) detecting said assay complexes by electrochemical detection.

### Claim 20 (depends on 19)

A method according to claim 19, further comprising digesting one strand of said amplicon using an exonuclease prior to step vii).

### Claim 21 (depends on 19)

A method according to claim 19, further comprising washing said detection electrodes prior to step ix).

### Claim 22

An apparatus for the detection of target analytes comprising:
a) an instrument bank comprising a plurality of biochip cartridge bays for insertion and analysis of a biochip cartridge, wherein each bay comprises:
i) a top bay comprising actuators for a liquid reagent module (LRM); and
ii) a bottom bay comprising electrical connections for an electrowetting electrode grid and detection electrodes; and
b) a base station comprising:
i) a central processing unit; and
ii) a user interface comprising a touch screen display having a plurality of bay icons, each icon uniquely corresponding to one of said plurality of bays.

### Claim 23 (depends on 22)

An apparatus according to claim 22, wherein when a biochip cartridge is inserted into one of said bays, said corresponding icon of said user interface is enlarged and/or exhibited.

### Claim 24 (depends on 22)

An apparatus according to claim 22, comprising a plurality of instrument banks.

### Claim 25 (depends on 1, 22)

A system for detecting a plurality of target analytes in a sample comprising:
a) a biochip cartridge according to claim 1; and
b) an apparatus according to claim 22.