# Electrokinetic paper diagnostic platform: 15-minute, quantitative nucleic acid amplification for viral pathogens in whole blood

> **NIH NIH R01** · UNIVERSITY OF WASHINGTON · 2020 · $484,674

## Abstract

Abstract
We propose to develop a nucleic acid based point-of-care diagnostic for quantifying viral load (VL) in whole
blood. The technology makes use of rapid RNA purification and reverse transcription recombinase polymerase
amplification (RT-RPA) on an electrokinetic paper device for quantitative detection of HIV-1 RNA in less than 15
minutes. This technology development is generally applicable to viral diagnostics in complex biological samples,
and here we focus on HIV-1 where no cost effective nucleic acid-based tests for point-of-care diagnostics are
available.
The HIV/AIDS epidemic is a major global health challenge, and is a leading cause of mortality and burden over
the last decade. Initiation of antiretroviral therapy (ART) can maintain durable viral suppression, however viral
load (VL) tests are needed to monitor the ongoing effectiveness of therapy, especially in prevention of mother to
child transmission of HIV. Currently, the optimal tests for measuring HIV VL rely on molecular amplification
methods that detect either HIV RNA or proviral DNA. A variety of commercial nucleic acid amplification
technologies (NAATs) assays are available for these purposes but their complexity, sensitive reagents, cost, and
hardware are prohibitively expensive and typically necessitate a fully functional centralized laboratory for their
use. The logistics around specimen collection, testing, and transport results in delayed diagnosis (from 9 days
to 5 months), diminished follow up with patients/guardians, and lost opportunity for life saving treatment.
We propose an electrokinetic paper based nucleic acid amplification assay that can quantify HIV VL from whole
blood in 15 minutes, with minimal user intervention, and no moving parts. Our innovative paper-based approach
combines rapid extraction of RNA by isotachophoresis (ITP) with RT-RPA amplification of target nucleic acids.
The diagnostic platform has integrated sample prep using a blood separation membrane, surfactant based virion
lysis, and ITP RNA purification. ITP extracts the nucleic acid targets from fractionated plasma while focusing the
RNA with RT-RPA reagents to accelerate the amplification reactions. The RPA reactions are quantified using
fluorescence intensity and an internal control reaction. A mobile phone is used to power and control the extraction
and reaction, measure the RPA reaction fluorescence, analyze the fluorescence data, and unambiguously report
the VL to the clinician and transmit to the cloud. The platform will provide quantification of HIV VL in 15 minutes,
for less than $10 per test, and minimal user intervention. The diagnostic will be validated over a wide range of
HIV subtypes and evaluated using clinical whole blood samples from HIV/AIDS patients.

## Key facts

- **NIH application ID:** 9960481
- **Project number:** 5R01EB022630-04
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Jonathan D Posner
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $484,674
- **Award type:** 5
- **Project period:** 2017-09-15 → 2023-07-29

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9960481

## Citation

> US National Institutes of Health, RePORTER application 9960481, Electrokinetic paper diagnostic platform: 15-minute, quantitative nucleic acid amplification for viral pathogens in whole blood (5R01EB022630-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/9960481. Licensed CC0.

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