Trends in Nanotechnology-based

diagnostics: An overview
(Dr. Pradip Patel)
IAFP, 18-20 May

Content

Why the need?

Current technologies and market data
Nanotech-based diagnostics: Examples of hand-held and
portable POC platforms
Bottlenecks and future prospects

Drivers 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

Industrial Microbiology Market

(courtesy of Tom Weschler, Strategic Consulting)

POC: $14.9 Bn Market Growing

(TSG & The Sharma Group, 2009)
at ~12% Annually
User needs are evolving, so no single platform likely to
address needs of all 4 POC user segments

$1.1Bn

Acute Care/
Hospital

TTR most important for critical care users

$4.1Bn

Physician Office
Lab (POL)

$9.4Bn

Self-test /
Consumer

Consumer DTX increasing tests must

be simple, low-cost & give quick results

$0.3Bn

Retail Clinic

Health clinic users seek fast, easy & lowcost for common, non-critical conditions

POL prefer results in <30m, expect high

performance & existing system integration

Current 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)

Dynamic 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)
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Frequently used Biosensor Principles in POC

Nanotech-based Diagnostics

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

Examples 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

Magnotech Hand-Held POC Platform

Courtesy of Jeroen Nieuwenhuis (Philips Healthcare Incubator)

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
Cartridge for blood Analysis is:
8 mm X 3 cm

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

10

Principle of Magnotech Assay

Courtesy of Jeroen Nieuwenhuis (Philips Healthcare Incubator)

Target capture

Magnetic attraction

Magnetic removal

Fast target
capture

Analyte
concentration

Effective bound/free
separation
11

Frustrated Total Internal Reflection

(FTIR) Readout Principle

Courtesy of Jeroen Nieuwenhuis (Healthcare Incubator)

12

Typical FTIR Signal Sensorgram

Courtesy of Jeroen Nieuwenhuis (Healthcare Incubator)

Signal decreases when beads bind to the surface due

to scattering and absorption of the evanescent field
13

Sandwich Assay: Cardiac Troponin I (TnI)

Courtesy of Jeroen Nieuwenhuis (Healthcare Incubator)

TnIdose re sponse
1 0 0 .0 0

)
%
(l
a
n
gi
S

1 0 .0 0

100% plasma
5 minutes
dry reagents

1 .0 0

0 .1 0
0 .0 1

0 .1

10

C o n cen tratio n(n g /m l)

14

14

Portable StatAnalyser: Core PCR-CE chip

(Courtesy of Maz Khorasani; Biolithic corp)

15

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)
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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!

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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

18

Schematic of a Typical Meat Homogenate

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

19

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

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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

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