nmed presentation

Development of Hand-held Instrument for STI Diagnostics

Professor.((W.(Balachandran((Bala)(Director((of(the(Centre(For(Electronic(Systems(Research(

School(of(Engineering(&(Design(Brunel(University(

emstwwb@brunel.ac.uk(

March 2013 1"

Enabling(and(TranslaGng(Advances(in(DiagnosGc(and(CommunicaGon(Technologies(to(Reduce(the(Burden(of(Sexually(

TransmiJed(InfecGons(

Self Testing Instrument for Sexually Transmitted Infections

Taking laboratory diagnosis into the field

Integrated System for POCT

3"

((((((Wireless(((((((Interface(

DNA(ExtracGon((&(PurificaGon((

Sample(collecGon( MicroFluidic(Network((

DNA((DetecGon(

Electronic(Control(System(

DNA((AmplificaGon(

Sample(concentraGon((&(cell(lysis(

The Team

Professor"Wamadeva"Balachandran"(Bala)"Principal(InvesGgator

Dr"Jeremy"Ahern"MicrofabricaGon Dr"Nada"Manivannan"

MulGphysics(Modelling Professor"Chris"Hudson"Electronic(Engineering(

Professor"Rob"Evans"

Biosciences(

Dr"Predraig"Slijpevic"""

Biosciences

Pascal"Craw"PhD(Student(

Biomedical(Engineering(

Branavan"Nehru"PhD(Student(

Paper(microfluidics(

Dr"Yanmeng"Xu"Printed(Electronics(

Sara"Chaychian"PhD(Student(

Electrical(Engineering(

Tosan"Ereku"PhD(Student(

Engineering(Design(

Dr"Krishna"Burugapalli""""""Biomedical(Engineering

Shavini"Wijesuriya"PhD(Student(

Engineering(Design(

Sana"Hussain"VisiGng(Scholar(Biosciences(

Sivanesan"Tulasidas""PhD(Student"Wireless(

CommunicaGon"""

Dr"Ruth"Mackay"BioMEMS/NEMS

Modular Research Platform

Electromagnets

Lysis

AmplificaGon

DetecGon

Sample(&(Reagents

Waste

Disposable(Cartridge

GPSRFID

Bluetooth( 3G(Mobile

USBWiFi

Microcontroller(

Power(Management(

Display(and(User(Interface

MagneGc

Electrochemical

OpGcal(

SPR

MEMS

Nanowire

Nucleic(Acid(Detector(

Pumps

Valve(actuators

Thermal(control

StandardisaGon

ConcentraGon(/(PurificaGon

Sample(preWtreatmentElectronic(System

Control(System(

CommunicaGon(

Interface

Valves

Sensors

Electromagnets

Microfluidic(Network(

Pathways

Sample Collection

•  Swab and urine •  4mL of urine •  100uL swab elute •  Simple design ‘Fool-

proof’ •  Direct integration to

extraction device •  Integrated lysis

Urine collection devices

FE Analysis to Inform Design

Streamline depiction of flow from inlets to device discharge orifice

CessaNonal" flow" of" urine" from" six"inlets"into"the"airQfilled"cavity"

DNA Extraction using Superparamagnetic Beads

8"

Ports

Coil recessCentral stub to Assist

the Coil Location

1 mm PMMA

Side Via to Port

500um PDMS

9 mm

Port Position

9 mm Coil Recess Diameter

Position of Chamber in PDMS Layer

12mm Pitch Circle for Ports

Chamber Cut-outPort Vias

8 mm

(a)

(b)

(c)

Microfluidic Chamber

Stainless Steel Port-ways

Polystyrene Collars

250mm Bore PVC Tubing

A photograph of microfabricated chamber with PVC tubing

Microfluidic Dispenser

DNA Extraction using Cationic Biopolymer

•  Novel membrane in development •  Cationic bioploymer membrane •  Reduces number of steps for DNA

extraction •  No toxic reagents •  Simple pH (5-9) change in aqueous

solutions •  2 reagents required •  Simple f low over device: no

centrifugation/active mixing

Two(DNA(extracGon(devices(with(embedded(biopolymer(membrane(

DNA Extraction Performance

0"

10"

20"

30"

40"

50"

60"

70"

80"

90"

100"

0" 0.1" 100"

Percen

tage(Recovery(((%

)(

Sample(ConcentraGon((ng/uL)((Salmon(sperm(DNA)(

Spin"Column"(Qiagen)"

Bioplymer"membrane"

Lab-in-a-Tube

Biopolymer and Lysis buffer will be integrated into the device to extract DNA

Cell Lysis and DNA Extraction"

0"

1"

2"

3"

4"

5"

6"

7"

8"

9"

10"

Standard"25µL""tube"rea

OnQchip"25µL"reacNon"

Final"D

NA"concen

traN

on"(u

g/mL)"

On-chip helicase dependent amplification

Isothermal Amplification •  Helicase dependent

amplification •  Single temperature (65�C) •  109 amplification power •  < 20minutes reaction time •  Can be used with real-time

fluorescence chemistries RealWGme(plot(of(HDA(reacGon(

Fluo

rescen

ce"

Time"(minutes)"0""""""""""""""""""""5"""""""""""""""""""""10""""""""""""""""""15"""""""""""""""""""20"""""""""""""""""""25"""""""""""""""""""30"""""""""""""""""""35""""""""""""""""""40"

NegaNve"control"

PosiNve"Control"

On-chip Amplification and Detection

Fluorescence"detecNon"on"microfluidic"chip"

0"

490nm"LED"

Amplified"Photodiode"

Emission"bandQpass"Filter""(530nm)"

OpNcal"Fibre"3mm"PMMA"

ReacNon"Chamber"

PMMA"Fluidic"Chip"

Finite"element"analysis"of"microfluidic"chip"to"characterise"thermal"properNes"

25µL"microfluidic"chip"

Planar Spiral Inductor for Inductance-based biosensor

Magnetic Bead-based DNA Detection

Silver"NPs;"a)1,"b)5"and"c)10µl"Hydrazine"in"0.1M"AOT/IPM"microemulsions"

Gold"NPs;"e)"1,"f)5"and"g)10µl"Hydrazine"in"0.1M"AOT/IPM"microemulsions"

a" b" c"

e" f" g"

Nano-particle/bead fabrication

Inductance Sensor: Simulation

(Circular(planar(coil(I(=(100(mA,((N(=(5((Maximum(electric(potenGal(=(160((mV(MagneGc(Flux(Density(=((4"Q"16(mT((

(Square(planar(coil(I(=(100(mA,((N(=(5((Maximum(electric(potenGal(=(110((mV(MagneGc(Flux(Density(=((4"Q"12(mT((

18"

Effect of dout on δL Effect of Beads Permeability µrB on δL

tc =2µm

tc =2µm

19"

Effect of Conductor Thickness tC on δL Effect of Thickness of Underlying Permalloy tp on δL

The Effect of Frequency on Sensor Output

Brass/Al"mould"for"a"detecNon"microfluidic"device"

DetecNon"device"with"automated"fluid"flow"and"electrodes"

Al"mould"for"a"fully"integrated"microfluidic"system"

Integrated"microfluidic"PDMS""device"

10mm"

10mm"

10mm"

Integrated Microfluidic Cartridges

Communication Design Strategy

Paper based microfluidics (µPADs)

Fabrication of µPADs

Wax"penetraNon:"comparison"of"printed"barriers"before"and"aher"curing"at"120oC"for"15"minutes"

Printed"barriers"of"500"µm"produced"fully"funcNonal"barriers."A"minimum"channel"width"of"~"300"

µm"is"achievable."

Printed"barriers"(Wax)"

Cured"barriers"(Wax)"

Xerox"ColorQubeTM"8570N"solid"ink"

Printer"

MulNplexing:"A"single"sample"effecNvely"

delivered"into"5"test"zones"

DNA"mobility"on"a"µPAD"

Inkjet"printed"silver"electrodes"(25"µm)"

DNA Detection on µPADs

0(s( 30(s( 90(s( W(1( W(2( 0s"–"A"blank"test"device"as"a"control"before"introducNon"of""""FITC"tagged"25mer"DNA"sample"(0.01nM).""

30s"–"DNA"sample"moving"within"the"hydrophilic"channel.""

90s"–"Further"movement"of"the"sample"into"the"waste"zone.""

W1"–"DNA"sample"gelng"washed"away"by"water"into"the"waste"zone.""

W2"–"Further"washing"of"the"DNA"by"water"into"the"waste"zone."

Water"as"control""

Blank"paper"as"control""

Serially"diluted"0.1pM"DNA""

Serially"diluted"1pM"DNA""

Serially"diluted"0.01nM"DNA""

Stock"DNA"soluNon"0.1nM"

All"above"pictures"are"obtained"through"the"BIOQRAD"Gel"DOCTM"XR+"system"and"the"associated"image"analysis"sohware"Image"LabTM."""

Handheld Device Development

The current handheld system development

Future GUI