Researchers at Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR) have developed a cutting-edge technology to enhance the accuracy of HIV diagnostics. Known as the GQ Topology-Targeted Reliable Conformational Polymorphism (GQ-RCP) platform, this novel diagnostic method leverages a fluorometric test to specifically detect G-Quadruplex (GQ) structures, a unique four-stranded DNA conformation in the HIV genome. This advancement promises to significantly reduce false positives in HIV testing while offering a reliable and efficient solution for early detection.
HIV-1, the retrovirus responsible for acquired immunodeficiency syndrome (AIDS), remains a major global health challenge. Conventional diagnostic tools, though widely used, have limitations, particularly in detecting early-stage infections. These methods often produce false positives due to cross-reactivity or fail to distinguish between nonspecific and target DNA sequences. Prolonged processing times and limited sensitivity further exacerbate these challenges.
Current nucleic acid-based diagnostics often rely on general DNA probes, which lack specificity and contribute to inaccurate results. Addressing this issue requires targeting unique nucleic acid sequences like G-Quadruplex (GQ) structures, which are characteristic of pathogenic genomes. By focusing on such noncanonical DNA conformations, researchers aim to revolutionize diagnostic assays and enhance reliability.
The GQ-RCP platform introduces a paradigm shift in molecular diagnostics by targeting unique DNA structures.
Dr. Thimmaiah Govindaraju, JNCASR
The GQ-RCP platform, developed by JNCASR scientists Sumon Pratihar, Vasudhar Bhat S.V., Krithi K. Bhagavath, and Thimmaiah Govindaraju, represents a significant innovation in molecular diagnostics. Initially designed for detecting SARS-CoV-2, this versatile platform has been successfully adapted for HIV detection, underscoring its modular potential.
The team demonstrated the reliable detection of HIV-derived GQ DNA by reverse transcription and amplification of a 176-nucleotide genomic segment. This process involves a pH-mediated, single-step transition of double-stranded DNA (dsDNA) into the GQ conformation, which is then detected using a benzobisthiazole-based fluorescent probe called TGS64. The study, published in Analytical Chemistry, highlights the remarkable selectivity of this method, setting the stage for a more precise and robust diagnostic platform.
“This innovation addresses a critical gap in HIV diagnostics by minimizing false positives,” noted the JNCASR research team. The platform’s ability to distinguish between nonspecific and target sequences positions it as a breakthrough in diagnostic technology.
Unlike most contemporary diagnostic assays, which rely on established detection principles, the GQ-RCP platform introduces a novel approach based on specific interactions between nucleic acids and small molecules. The following features set it apart:
Targeted Specificity: The platform’s molecular probes recognize GQ structures with exceptional precision, reducing false positives.
Fluorometric Detection: The use of fluorescent probes like TGS64 ensures high sensitivity and selectivity.
Integration Potential: GQ-RCP can be incorporated into existing nucleic acid-based diagnostic systems to enhance their reliability.
By addressing limitations in amplification-based techniques, this platform offers unambiguous detection of noncanonical DNA conformations, a critical advancement in molecular diagnostics.
This innovation addresses a critical gap in HIV diagnostics by minimizing false positives.
JNCASR Research Team
The versatility of the GQ-RCP platform extends beyond HIV detection. Its ability to accurately identify DNA/RNA-based pathogens makes it suitable for diagnosing various infectious diseases, including bacterial and viral infections. The platform’s adaptability and modular design ensure its potential for widespread adoption in clinical settings.
Dr. Thimmaiah Govindaraju from JNCASR emphasized, “The GQ-RCP platform introduces a paradigm shift in molecular diagnostics by targeting unique DNA structures.” This approach not only addresses existing diagnostic challenges but also opens new avenues for precision-based pathogen detection.
The development of the GQ-RCP platform marks a significant step forward in the fight against HIV/AIDS. By leveraging noncanonical DNA conformations, researchers have created a diagnostic tool that overcomes the limitations of current methods, offering a reliable and efficient alternative. With its potential for integration into existing systems, the GQ-RCP platform promises to revolutionize pathogen detection and strengthen global health efforts.
Reference:
1. Sumon Pratihar, Vasudhar Bhat S V, Krithi K. Bhagavath, and Thimmaiah Govindaraju. Unambiguous Detection of LTR-III G-Quadruplex in the HIV Genome Using a Tailored Fluorogenic Probe-based Assay. Analytical Chemistry 2024 96 (40), 15834-15839. DOI: 10.1021/acs.analchem.4c03374
(Input from various sources)
(Rehash/Yash Kamble/MSM)
