Desenvolvimento de reagente para extração rápida de ácidos nucleicos e kit multiplex para detecção de variantes do SARS-CoV-2 por RT-qPCR

Abstract

Since the beginning of 2020, humanity has faced the challenges imposed by the COVID-19 pandemic, a respiratory disease caused by the SARS-CoV-2 virus. The global response to contain this pandemic focused, among other actions, on the development of accurate diagnostics, effective vaccines, and the search for appropriate medications. However, significant obstacles were encountered in the field of diagnostics due to the shortage of materials for sample collection, processing, and analysis through RT-qPCR, a key technique in virus detection. One of the main challenges faced was the delay and high cost of nucleic acid extraction procedures, which hindered efficient diagnostic efforts. Furthermore, the complex procedures involved in sample preparation increased the risk of contamination and delayed the diagnostic process. In addition, the emergence of new virus variants highlighted the need for epidemiological surveillance to contain new waves and the importance of detection kits capable of supporting monitoring actions. In this context, the present work aimed to develop a rapid nucleic acid extraction reagent to speed up and make RT-qPCR diagnosis more accessible. Moreover, the study proposed the development of a multiplex kit for the detection of SARS-CoV-2 variants using this technique. The results obtained demonstrated that the developed reagent enabled reliable analyses through qualitative PCR. This reagent not only proved to be effective but also advantageous in reducing costs associated with laboratory supplies and plastic materials, in addition to minimizing the risk of contamination. The multiplex kit for the detection of SARS-CoV-2 variants showed efficacy in differentiating between variants, demonstrating that RT-qPCR can be an essential tool in monitoring the virus's evolution. With the successful implementation of this reagent and the multiplex kit, it is expected that the diagnostic process for many diseases will be expedited and that the emergence of new variants will be better tracked. This technological advancement represents a milestone in the fight against the pandemic, offering a viable and efficient solution for the rapid and reliable detection of the virus. Furthermore, its application may significantly contribute to controlling the spread of the disease and strengthening global public health.