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A microfluidic based point of care testing device for measuring urine albumin using a novel organic dye

Primary Information

Domain

Healthcare

Project No.

6060

Sanction and Project Initiation

Sanction No: 3-18/2015-TS-TS.1

Sanction Date: 29/11/2016

Project Initiation date: 08/02/2017

Project Duration: 36

Partner Ministry/Agency/Industry

Indian Council of Medical Research Emware Technologies (India) Pvt. Ltd.

Role of partner:Indian Council of MEdical Research will provide funds to carry out prototype development and also provide guidelines to carry out clinical trials. Emware Technologies (India) Pvt. Ltd will help us develop the PCB of the final product.

Support from partner:The partner industry will help us in the translation from field level prototype to product translation and later on in the commercialization process also.

Principal Investigator

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Dr. Shubhajit Roy ChowdhuryPrimary
Indian Institute of Technology Mandi

Host Institute

Co-PIs

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Dr. Subrata Ghosh
Indian Institute of Technology Mandi

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Dr. Prosenjit Mondal
Indian Institute of Technology Mandi

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Dr. Chander Singh
Indian Institute of Technology Mandi

Scope and Objectives

1. Estimating the exact concentration of albuminin urine through the development of a low cost point of care testing device using a novel NIR based lumino-material composed assay system.
2. Obtaining exact quantification of urinary albumin without any disturbance arising from the auto-fluorescence of the bio-molecule present in urine.
3. Prognosticating a wide variety of nephritic diseases by a long range linear correlation between the fluorescent signal of probe and the amount of urinary albumin.
4. Clinical trial of the proposed device to increase the class and effectiveness of the system.

Deliverables

The project is aimed at delivering a low cost handheld portable device to estimate the albumin concentration in urine particularly when albumin is very less in urine. The detection will be done by measuring fluorescence intensity of the probe. This fluorescent signaling mechanism is considered as one of the most sensitive methods, therefore this NIR based quantification measurements will avoid the auto fluorescence of biomolecules exist in urine.

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Videos

https://www.youtube.com/watch?v=GhnFAunvbwg&feature=youtu.be

Scientific Output

The proposed device has correctly estimated albumin concentration in urine with as low concentration as 3.3ug/dL which in terms of normality is 50 nM concentration. This is in fact, the least concentration correctly detected so far. In order to estimate albumin in urine and to ensure that there is no interference coming from biospecies in urine, we developed a dye that binds with albumin in urine. The excitation of the dye+urine mixture happens at 740 nm and emission takes place at 804 nm wavelengths. The dye is highly photostable and nontoxic new NIR probe (CyG) for detection of albumin in different biofluids. Also, NIR dyes have gained popularity as compared to visible dyes due to minimized tissue autofluorescence, deep tissue imaging capability through efficient noninvasive penetration, reduced photon scattering and high contrast image.

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Results and outcome till date

Linearity in the behavior of the device has been achieved with up to 50nM concentration. The results that we have got from our generation 1 prototype device is very reproducible and linear, which indicates that our proof of concept is vindicated and we wish to proceed to generation 2 prototype device soon.

Societal benefit and impact anticipated

The concentration of albumin in various biofluids particularly in urine could be used as potential indicator of early stage of various human health disorders that include kidney damage, diabetes, and so on. Moreover, urine dip-stick device is already available in market for semi- quantitative detection of albumin in urine, but some of its limitation restricted it from measuring the exact amount of albumin present in urine. Those limitations are 1. Microalbuminuria cannot be detected by using urine strip model. The detection limit of urine stick procedure for detecting albumin in urine is minimum 150 mg /L which is quiet higher than the albumin level present in urine during microalbuminuria. 2. It also provides limited information about the urine as it is a kind of qualitative test and not a quantitative test (for example, it does not give a precise measure of the quantity of abnormality) 3. Color of the urine affects the result produced by urine strip model. 4. The absorption of the dye used in urine strip model lies in 480-550 nm region which can be disturbed by the presence of other biomolecules in urine sample. 5. High salt level will lower the result. 6. A false positive result is seen on alkaline /highly buffered urine samples, this limitations encouraged us to develop a low cost device for estimation of albumin in urine using a spectrophotometric approach. The current project involves development of a microfluidic device that accepts urine and dye through two separate channels and then entered into a reaction site where LED and photodetectors will be placed. The channels will be optical fiber channels to aid in the spectrophotometry. The collector current of the photodetector will be amplified using a band pass amplifier to remove out of band noise and then digitized and displayed using an LCD.The diagnostic assay also needs to be validated. We shall also follow the international guideline to validate our proposed diagnostic assay tool. Our developed set up can be used for detecting other analytes by changing bio-molecule specific different organic dye to make the instrument more versatile.

Next steps

Currently we are in the process of translation from lab level prototype to field level prototype. Once the field level prototype is ready we shall undergo extensive clinical trials o a pilot basis at the Medical Unit of IIT Mandi.

Publications and reports

Will be communicated soon.

Patents

S. Roy Chowdhury, S. Ghosh, P. Mondal, Y. Arora, B. Biswas, S. Mukherjee, V. Bedi, "A System for detecting biological molecule and method of using the same", Indian Patent Application no. 201811047739 dated December 17, 2018.

Scholars and Project Staff

1. Mr. Soham Mukherjee - Project Associate : Working since June 2018. 2. Ms. Bidisha Biswas - Project Associate: Working since June 2017.

Challenges faced

The following are the difficulties faced during development: 1. Alignment of the LEDs and photodetector with respect to the sample. 2. Improvement of SNR of the output using low power LEDs. 3. Estimation of albumin in urine when other bio-species are present. 4. Temperature drift compensation of the system 5. Ensuring no other radiation interferes with the device.

Financial Information

  • Total sanction: Rs. 7320000

  • Amount received: Rs. 4780000

  • Amount utilised for Equipment: Rs. 1160000

  • Amount utilised for Manpower: Rs. 780000

  • Amount utilised for Consumables: Rs. 1280000

  • Amount utilised for Contingency: Rs. 900000

  • Amount utilised for Travel: Rs. 130000

  • Amount utilised for Other Expenses: 0

  • Amount utilised for Overheads: Rs. 780000

Equipment and facilities

AFG-1062 Arbitrary waveform generator. Tektronix make. 2 channel, 60 MHz bandwidth, 300 Msa/s, 1M sampling points.