New Technology Developed for Precise Detection of HIV Genome Using Tailored Fluorogenic Tests

Researchers have developed an innovative technology to enhance the detection of the HIV genome, specifically targeting the G-Quadruplex (GQ) structure, a unique four-stranded DNA formation found in the virus. This new fluorometric test offers a highly reliable diagnostic method, significantly reducing the risk of false positives in HIV detection.

HIV-1, the retrovirus responsible for AIDS, continues to pose a global health threat. Existing HIV diagnostic methods can sometimes fail to detect early infections or result in false positives due to cross-reactivity. Additionally, other clinical methods for early detection are limited by reduced sensitivity and long processing times. Current nucleic acid-based tests are prone to false positives because general DNA probes fail to distinguish between nonspecific and target amplicons. Therefore, identifying and targeting specific nucleic acid sequences is crucial in minimizing these errors. By focusing on unique nucleic acid structures like the GQ formation, researchers have made a significant advancement in creating precise diagnostic tests.

A team from the Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR), an institution under the Department of Science and Technology, developed the GQ Topology-Targeted Reliable Conformational Polymorphism (GQ-RCP) platform. Originally designed for detecting pathogens like SARS-CoV-2, the versatile platform has now been successfully adapted for HIV detection, showcasing its modular capabilities.

Researchers Sumon Pratihar, Vasudhar Bhat S V, Krithi K. Bhagavath, and Thimmaiah Govindaraju demonstrated the detection of HIV-derived GQ DNA using a reverse transcription and amplification technique that targets a 176-nucleotide segment of the virus’s genome. A key part of their study, published in Analytical Chemistry, was the development of a pH-mediated, single-step process that converts double-stranded DNA into the GQ conformation, the target for detection. This transition was observed with exceptional selectivity using a benzobisthiazole-based fluorescent probe (TGS64).

Unlike most other recent diagnostic advancements, this research introduces a novel platform based on specific interactions between nucleic acids and small molecules, offering a new approach to pathogen detection. The GQ-RCP method promises to reduce the challenge of false positives in existing amplification-based techniques by providing sequence-specific recognition and clear detection of the target GQ structure.

This new detection platform can be integrated with current nucleic acid-based diagnostics, improving their reliability. It is expected to be adaptable for detecting a wide range of DNA and RNA-based pathogens, including other viruses and bacteria. This breakthrough could significantly enhance the accuracy and efficiency of diagnostic tests worldwide.