A group of scientists has developed a new technology platform for fluorometric detection of diseases such as viruses using measurements of fluorescent light emitted, and the new technology's potential has been proved using SARS-CoV-2 detection.
Other DNA/RNA diseases, such as HIV, influenza, HCV, Zika, Ebola, bacteria, and other mutating/evolving pathogens, can be detected using this technology platform.
The current Covid-19 epidemic, which is caused by SARS-CoV-2, is wreaking havoc on all facets of our lives, demonstrating how viruses are a huge global threat to human health. The exceptional rate of RNA virus transmission has prompted speedy and accurate diagnosis to assist contact tracing (and hence prevent the virus from spreading).
Scientists from the Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR), an autonomous institute of the Indian Ministry of Science and Technology, and scientists from the Indian Institute of Science (IISc) have demonstrated a noncanonical nucleic acid-based G-quadruplex (GQ) topology targeted reliable conformational polymorphism (GQ-RCP) platform to diagnose Covid-19 clinical samples.
"Based on a novel platform GQ-RCP, the present work demonstrated the first GQ-targeted diagnostic platform for SARS-CoV-2 in clinical samples," according to a release from the Ministry of Science and Technology. "This molecular detection platform can be integrated into field-deployable isothermal amplification assays with more reliability and sequence specificity."
This work was recently published in the journal ACS Sensors by Sumon Pratihar, Ragini Agrawal, Virender Kumar Pal, Amit Singh, and Thimmaiah Govindaraju (corresponding author).
To achieve stable and dependable noncanonical DNA/RNA targets, the platform places a larger emphasis on deciphering and systematic characterization of a unique set of interactions in nucleic acids. The RCP-based target validation methodology is a generic and modular method for developing noncanonical nucleic acid-targeted diagnostic tools for a wide range of diseases, including bacteria and DNA/RNA viruses.
For reliable detection of SARS-CoV-2, RT-q-PCR has been the gold standard (Covid-19). Techniques like RT RPA and RT-LAMP, which use general-purpose DNA sensing probes, are among the most recent innovations in the nucleic acid-targeted diagnosis of SARS-CoV-2. As a result of unbiased identification of nonspecific amplification products, the likelihood of false-positive outcomes increases. Recognizing distinct DNA secondary conformations could be a potential method for obtaining accurate readouts.
The researchers discovered and characterized a new G-quadruplex-based target generated from SARS-30 CoV-2's kb (kilobytes) genomic landscape for SARS-CoV-2 detection. Unlike prior reliable diagnostic tests that have reused existing fundamental concepts, this work uses small-molecule fluorophores (microscopic molecules) to target a unique, atypical structure exclusive to the SARS-CoV-2 sequence, according to the release.
The researchers created GQ topological tailored detection of SARS-CoV-2 derived DNA in clinical samples collected following reverse transcription and amplification from genomic RNA. The pH-triggered easy transformation of amplified double-stranded DNA into stable GQs, which forms the target for detection with amazing selectivity using a developed fluorescent dye that is a benzobisthiazole-based fluorescent dye, was used to enable GQ targeted detection.
"As a result, our study demonstrates a reliable technique for fluorogenic organic molecule-based GQ-RCP platform to diagnose Covid-19 clinical samples and is the first practical demonstration of it," the researchers wrote.
"We've shown how to use rational molecular probe tailoring to obtain unambiguous target recognition and increased detection reliability in less time, without the need for an expensive RT-q-PCR device. This RCP-based platform is very versatile and may be used to identify a wide range of DNA/RNA-based diseases, such as bacteria and viruses like HIV, Influenza, and HCV "Govindaraju stated.
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