The advancement of antiretroviral treatment (ART) has significantly reduced the HIV mortality and morbidity by reducing the viral load (VL) to undetectable levels. Recently revised WHO guidelines strongly recommend routine VL testing to monitor ART adherence and minimize failure. To this end, HIV self-testing, a process in which individual who wants to know HIV status collects a specimen, performs a test and interprets the result in private, has become an empowering approach. While nucleic acid testing (NAT) is readily available in centralized laboratories for viral load quantification, its availability in self-testing has not been demonstrated due to sample processing and assay complexity. This project aims to develop a quantitative test on an ultra-compact USB device to detect viral rebound that is simple enough for laypersons to test themselves in the United States. The R61 phase of the project will develop the whole blood-based test that can quantitatively assess the presence of HIV-1 RNA at concentrations as low as 1000 copies/ml, threshold recommended by WHO for determining treatment failure.
In aim 1, we will develop a disposable microfluidic chip for streamlined and automated plasma separation and viral RNA extraction from whole blood.
In aim 2, we will optimize the HIV-1 RT-LAMP assay and explore the minimum copy number sensitivity.
In aim 3, we will integrate the quantitative USB analyzer hardware and develop software for easy and robust operation.
In aim 4, we will validate the prototyped test in the BSL-2 lab using HIV-1 plasma samples spiked into whole blood. The R33 phase of the project will assess the test performance, usability and stakeholder needs within the HIV Comprehensive Care Program at Penn State Hershey Medical Center.
In aim 5, the performance of the proposed test on HIV-infected patients will be benchmarked with standard laboratory methods.
In aim 6, we will evaluate the user?s experience, attitude and perception of HIV VL self-testing. Through innovations in microfluidic chips and the USB analyzer, we anticipate the test would be able to quantify HIV-1 VL as low as 1000 copies/ml directly from 100 l of finger prick blood. The potential impact of this project is very high. If this proof-of-concept project is successful, it has the potential to significantly enhance the treatment outcomes for individuals of HIV under therapy. The potential transformative capacity warrants the challenges associated with this project.
HIV self-testing would benefit both high-risk populations unaware of their infection and individuals aware of their infection and under treatment. This project seeks to develop and validate an easy-to-use quantitative HIV self- testing device to allow lay individuals to perform HIV viral load measurement. Increasing access to routine viral load testing can significantly improve the health outcomes of people living with HIV and help to prevent the onward transmission of HIV towards ending AIDS as a public health threat.
