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PAPER-BASED MICROFLUIDIC POINT-OF-CARE DIAGNOSTIC DEVICES

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Transcript of PAPER-BASED MICROFLUIDIC POINT-OF-CARE DIAGNOSTIC DEVICES

Paper-Based Enzyme-Linked Immunosorbent Assay (P-ELISA)

The main advantages of a P-ELISA are –
It is cheaper and the results can be analysed using desktop scanner
The entire assay can be performed within an hour.
Requires less than 20µL of sample for analysis.

The main disadvantage is that it is less sensitive.

* The rabbit and goat can be replaced by other animals.
PAPER-BASED MICROFLUIDIC POINT-OF-CARE
DIAGNOSTIC DEVICES USED TO ANALYSE
BIOLOGICAL SAMPLES

Ms. M. B. P. Wickramaratne
10407
Presentation of Seminar
Department of Chemistry, University of Colombo

May 07th 2014
`
What are Microfluidic Point-of-Care Diagnostic Devices
Microfluidic Point-of-care (POC) diagnostic devices are devices which are capable of detecting constituents of a given sample, on-site, and producing results that can be used for an analysis immediately, by utilising only a minute amount of sample.
These devices can be used in the fields of immunoassays, food safety, veterinary medicine, environmental monitoring, bioterrorism, urine analysis etc.

Why are paper-based POC devices so attractive?

Cheap
Ubiquitous
Light-weight
Can be disposed of by incineration, so doesn’t produce biohazardous waste.
Can be produced on a mass scale
Portable
In the year 2004, the World Health Organisation (WHO) brought forth the concept “ASSURED”. This concept provides some guidelines according to which POC devices should be made so that they will be better suited for use in developing countries and in emergencies.

The “ASSURED” Concept



“ASSURED”

A
ffordable - Paper is cheap
S
ensitive - Can detect even small
concentrations
S
pecific - Detects the specific analyte only.
U
ser-friendly - Requires minimum user manipulation.
R
apid and robust - Gives results immediately and is
durable.
E
quipment-free - Requires no external pump as sam
ple

flows through device by capillary
action.
D
elivered - Should be portable to the
required location.

Applications of POC devices

As mentioned above POC devices can be used in a number of fields, but this seminar will focus only on their applications in analysing biological fluids.
Enzyme-Linked Immunosorbent Assay (ELISA)

ELISAs are immunoassays which are frequently performed to diagnose infectious diseases.

An ELISA is usually performed using a microtiter plate and they are quantified using a plate reader.

Advantages
Sensitive
Specific
Suitable for high-throughput assays
Disadvantages
Both the microtiter plate and the plate reader is very costly (about $ 20,000).
Each step of the assay takes over one hour.
Requires large volumes (20-200 µL) of analyte and reagents.

Source -
http://gmwgroup.harvard.edu/pubs/pdf/1086.pdf

Source - http://www.elisa-antibody.com/ELISA-Introduction/elisa-device

Source - http://gmwgroup.harvard.edu/pubs/pdf/1086.pdf

Source - http://gmwgroup.harvard.edu/pubs/pdf/1086.pdf

Point-of-care Diagnostics for Noncommunicable Diseases

In the past few decades a significant growth in the prevalence of NCDs has been observed around the world. However about 80% of the reported NCDs are from the developing countries and since 2001, cardiovascular diseases and cancer have been the leading causes of mortality in developing nations.
Proposed Experimental Design
Source - http://lmrt.mit.edu/publications/2014/Warren_PNAS_earlyed_SI_2014.pdf

Experiment

Experiment was conducted using mice. The representative diseases considered were thrombosis and colorectal cancer (CRC).

Method –

1) Poly(ethylene glycol)-coated iron oxide NWs were conjugated to
20-30 substrate-reporter peptides, per NW particle.
2)These were injected to mice diseased with thrombosis and allowed
to circulate in the blood stream.
3)Urine samples were collected and analysed using a lateral flow
assay (LFA).

Designing a Lateral Flow Assay

The LFA was designed to detect the reporters using a sandwich complex. Hence the reporters were specially designed for this purpose. This reporter acts as an antigen.

The linker was Glutamate-fibrinopeptide B (GluFib)


Source - http://lmrt.mit.edu/publications/2014/Warren_PNAS_earlyed_SI_2014.pdf

Source - http://lmrt.mit.edu/publications/2014/Warren_PNAS_earlyed_SI_2014.pdf

Given above are just two examples to show the applications of paper-based POC devices. However quite number of devices have been made to detect other constituents in blood, sweat, urine etc. An ideal POC device would be one that gives a colour change that is easily detectable even to the naked eye, in the presence of the analyte.


Perhaps in the future we will be able to conduct blood tests at the comfort of our homes !!!!

References

Vella, S. J., Beattie, P., Cademartiri, R., Laromaine, A., Martinez, A. W., Phillips, S. T., Mirica, K. A., and Whitesides, G. M. (2012) Measuring markers of liver function using a micropatterned paper device designed for blood from a fingerstick. Anal. Chem. 84, 2883–91.

Martinez, A. W., Phillips, S. T., Carrilho, E., Thomas, S. W., Sindi, H., and Whitesides, G. M. (2008) Simple telemedicine for developing regions: camera phones and paper-based microfluidic devices for real-time, off-site diagnosis. Anal. Chem. 80, 3699–707.

Yetisen, A. K., Akram, M. S., and Lowe, C. R. (2013) Paper-based microfluidic point-of-care diagnostic devices. Lab Chip 13, 2210–51.

Cheng, C.-M., Martinez, A. W., Gong, J., Mace, C. R., Phillips, S. T., Carrilho, E., Mirica, K. A., and Whitesides, G. M. (2010) Paper-based ELISA. Angew. Chem. Int. Ed. Engl. 49, 4771–4.

Warren, A. D., Kwong, G. A., Wood, D. K., Lin, K. Y., and Bhatia, S. N. (2014) Point-of-care diagnostics for noncommunicable diseases using synthetic urinary biomarkers and paper microfluidics. Proc. Natl. Acad. Sci. 111, 3671–3676.


Thank you !!!

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