Advancing Integrated Nucleic Acid Testing for HIV Self-Testing and Personalized Viral Load Monitoring
Restricted (Penn State Only)
- Author:
- Liu, Tianyi
- Graduate Program:
- Electrical Engineering
- Degree:
- Doctor of Philosophy
- Document Type:
- Dissertation
- Date of Defense:
- August 13, 2024
- Committee Members:
- Madhavan Swaminathan, Program Head/Chair
Zhiwen Liu, Major Field Member
Mehdi Kiani, Major Field Member
Weihua Guan, Chair & Dissertation Advisor
Cunjiang Yu, Outside Unit & Field Member - Keywords:
- Human Immunodeficiency Viruses
Viral Load
Self-Testing
Nucleic Acid Testing
Reverse transcription polymerase chain reaction
Quantification - Abstract:
- The Human Immunodeficiency Virus (HIV) remains a significant global public health challenge, particularly in resource-limited settings where access to timely and accurate viral load monitoring is constrained. This dissertation presents the development of a portable, user-friendly device designed for quantitative HIV detection from finger-prick blood samples, aiming to address the critical need for accessible HIV viral load testing in these regions. The developed device integrates advanced nucleic acid testing (NAT) techniques, such as reverse transcription loop-mediated isothermal amplification (RT-LAMP) and reverse transcription polymerase chain reaction (RT-PCR), into a compact, portable format. This innovation facilitates a seamless 'sample-to-answer' solution, enabling self-testing with rapid and accurate results without the need for extensive laboratory infrastructure or professional expertise. This dissertation focuses on the development of a portable, user-friendly device for quantitative HIV detection from finger-prick blood samples, tailored for use in resource-limited settings. The research encompasses the following key advancements: 1. NAT-on-USB Device: The creation of a fully integrated NAT device for HIV self-testing. This device features a microfluidic reagent cartridge and an ultra-compact analyzer interfaced via USB. It simplifies the testing process, requiring only minimal user interaction, and achieves a detection limit of 214 viral RNA copies/mL within 60 minutes. 2. Quantitative Detection Enhancements: Development of quantitative detection capabilities, including a disposable plasma separation card and a syringe-based RNA extraction module. These enhancements ensure efficient plasma separation and viral recovery, achieving 80% and 86% efficiency, respectively. Additionally, a smartphone interface for real-time semiquantitative RT-LAMP testing was implemented, maintaining stable performance over 16 weeks. 3. Multiplex PCR for HIV-1 and HIV-2: Advancing the device to perform multiplex RT-PCR for simultaneous detection of HIV-1 and HIV-2 viral loads. This portable PCR analyzer, designed for resource-limited settings, demonstrates high sensitivity and specificity, confirming its reliability and effectiveness. 4. Simultaneous Detection of HIV and HCV: Refinement of the multiplex PCR device to allow rapid self-testing for both HIV and Hepatitis C Virus (HCV) viral loads. The battery-operated system completes PCR amplification in 45 minutes and shows high accuracy and reliability, comparable to benchtop PCR systems. These innovations represent significant progress in making HIV viral load testing more accessible and practical, particularly in high-prevalence regions with limited healthcare resources. The developed device offers a rapid, accurate, and user-friendly solution for self-testing, which is crucial for improving HIV management and treatment outcomes globally.