Trends in Nanotechnology-based diagnostics: An overview

(Dr. Pradip Patel)IAFP, 18-20 May

2Content

Why the need?

Current technologies and market data

Nanotech-based diagnostics: Examples of hand-held and portable POC platforms

Bottlenecks and future prospects

3Drivers of Diagnostic Technology Development

Improve quality of patient care: Rapid therapeutic decisions based on paradigm shift from central lab to testing at Point-Of-Care (POC).

Food-borne illness

is a global issue. According to CDC&P (USA):~76 million cases annually; ~5000 deaths a year direct costs

$46 billion (lost productivity/health care)

Food quality/safety legislation

(e.g. EC Regulation 2073/2005 on microbiological criteria for foodstuffs linked with HACCP)

Food trade

(global competition)

Companies Internal food safety strategy

(HACCP, product right-first-time, due diligence, marketing)

Conventional methods

too time-consuming and labour-intensive

4Industrial Microbiology Market (courtesy of Tom Weschler, Strategic Consulting)

5TTR most important for critical care usersAcute Care/ Hospital

$1.1Bn

User needs are evolving, so no single platform likely to address needs of all 4 POC user segments

$4.1Bn

$9.4Bn

$0.3Bn

Physician Office Lab (POL)

Self-test / Consumer

Retail Clinic

POL prefer results in

6Current Microbiology Tests

Culture-based: media (Petri dishes, bottles, bags, tubes.)

Immunoassays: Manual/automated (ELISAs, lateral flow assays.)

Molecular amplification assays: Manual/automated platforms (RT-PCR, qRT-PCR, NASBA.)

Identification: Manual/automated ID systems (API, ATB, Vitek 2)

7Dynamic Industrial Needs!

Although current generation technologies adequately meet the needs of highly varied customer base (central & contract food and clinical labs, independent labs..), user requirements are continually changing:

Even lower analytical costs (often testing is viewed as a cost!)

Higher automation & integration with existing workflow

Reduced TTR (preferably results in few hours NOT days!)

No compromise on performance

High throughput with multiplex capability

Fit-for-purpose portable systems

Etc..

(Over the past decade, significant developments in nanotechnology, microfabrication (microfluidics, LOC devices), bioinformatics and connectivity have paved the way for technological developments that are expected to meet the challenges posed by the dynamic industrial needs)

8Frequently used Biosensor Principles in POC Nanotech-based Diagnostics

From: Luppa

et al., 2011. DOI: 10.1016/j.trac.2011.01.019

9Examples of POC Platforms

Hand-Held POC device (e.g. Philips Magnotech

biosensor)

Immunological principle

Nanomagnetic bead-based

Transduction mode: FTIR

Portable device (e.g. Biolithic StatAnalyser)

PCR-coupled CE chip

Detection: LIF

10

Magnotech Hand-Held POC Platform

Cartridge

for blood Analysis

is:8 mm X 3 cm

(Philips and bioMerieux are in partnership to develop POC devices for the IVD sector)

Quantitative detection ( nMpMand lower ) from a small droplet ( 15-40 L

) of whole blood

within minutes with

high reliability

multiplexed in a cost-effective cartridge with

a compact analyser

Courtesy of Jeroen Nieuwenhuis (Philips Healthcare Incubator)

11

Courtesy of Jeroen Nieuwenhuis (Philips Healthcare Incubator)

Principle of Magnotech Assay

Target capture

Fast target capture

Magnetic attraction

Analyte concentration

Magnetic removal

Effective bound/free separation

12

Frustrated Total Internal Reflection (FTIR) Readout Principle

Courtesy of Jeroen Nieuwenhuis (Healthcare Incubator)

13

Typical FTIR Signal Sensorgram

Signal decreases when beads bind to the surface due to scattering and absorption of the evanescent field

Courtesy of Jeroen Nieuwenhuis (Healthcare Incubator)

1414

Sandwich Assay: Cardiac Troponin I (TnI)

100% plasma 5 minutes dry reagents

0 .10

1 .00

10 .00

100 .00

0 .01 0 .1 1 10

Signal(%)

Concentrat ion(ng/m l)

TnIdose response

Courtesy of Jeroen Nieuwenhuis (Healthcare Incubator)

PresenterPresentation NotesSignificance: Good linear range of almost 3 orders of magnitudeSignificance: the LOD is well below 1 pM (1 pM = 0.0259 ng/ml)

15

Portable StatAnalyser: Core PCR-CE chip(Courtesy of Maz Khorasani; Biolithic corp)

16

Bakulovirus Detection by PCR-CE Chip (Kaigala

et al (2010), Analyst, 135, 1606)

The PCR product peak (299 bps) represents the presence of BK virus in a sample LOD quoted is about 20pg/uL

(similar to agarose

gel/ethidium

bromide method)

17

StatAnalyser: Essentially a LOC Toolkit(Courtesy of Maz Khorasani; Biolithic corp)

Sold as modular portable toolkit for POC application, NOT

as final assays

Performance claim:equivalent to other

expensive

commercial systems

The entire system in its current form is shoebox-sized Total component cost of ca. $600!

18

So, what are the Bottlenecks?

Sample matrix complexity

Background components

Compositional variability

Potential interference from

non-target flora

PCR

iInhibitors

Lengthy analysis

times due to inevitable

cultural growth steps

Thus, results

are generally only

of retrospective

value

19

Schematic of a Typical Meat Homogenate

Patel(2000). J.Rap.Meths. & Automa. Microbiol., 8, 227-248

20

Dielectrophoresis (DEP) : Selective Isolation of Live from Dead Human Leukemia Cells

No electric

field: Dead (red) and live (blue) cells

moving

right to left

.

Electric field

(30 s):

live cells

are trapped(+ve

DEP); dead

cells

pass

by

Power supply

off: live cells

released

From: Shafiee et al., 2010. DOI: 10.1039/b920590j

21

Prospects

Developments

in capture/concentration as alternatives/adjuncts

to cultural growth

Improved

high

productivity/selectivity

media

Recent

technological

developments

in microfabrication, microfluidics, LOC, multiplexing

and bioinformatics

and IT communication devices

Overall, integration

of the above

areas are expected

to deliver

the next

generation

diagnostic technologies

Slide Number 1Content Drivers of Diagnostic Technology DevelopmentIndustrial Microbiology Market (courtesy of Tom Weschler, Strategic Consulting)User needs are evolving, so no single platform likely to address needs of all 4 POC user segmentsCurrent Microbiology TestsDynamic Industrial Needs!Frequently used Biosensor Principles in POC Nanotech-based Diagnostics Examples of POC Platforms Magnotech Hand-Held POC PlatformPrinciple of Magnotech AssayFrustrated Total Internal Reflection (FTIR) Readout PrincipleTypical FTIR Signal SensorgramSandwich Assay: Cardiac Troponin I (TnI)Slide Number 15Slide Number 16Slide Number 17So, what are the Bottlenecks? Schematic of a Typical Meat Homogenate Dielectrophoresis (DEP) : Selective Isolation of Live from Dead Human Leukemia Cells Prospects