Método Multiplex RT-qPCR para detección de virus que causan infecciones respiratorias agudas graves en el Ecuador

Abstract

Severe acute respiratory infections (SARI) cause the death of nearly 4 million people annually, making it the sixth leading cause of mortality in all age groups and the leading cause of death in children under 5 years of age. On the other hand, Ecuador's dependence on foreign technology to implement diagnostic tests implies high costs and a long import time. Considering the importance of epidemiological surveillance and the benefits of a rapid and safe diagnosis, the need arises to develop locally a method for the detection of viruses that cause SARI in Ecuador. The objective of this project is to implement a local multiplex diagnosis of respiratory viruses using the RT-qPCR technique for its application in public health. To this end, primers and probes were designed to detect Influenza A/B, RSV and SARS-CoV-2; then, optimization tests were performed in individual and multiplex reactions to finally validate the test with positive samples. The multiplex PCR simultaneously detected the 3 targets in a single reaction tube and the results with positive samples coincided 100% with the expected diagnosis, demonstrating that it is feasible to use it as a diagnostic method. The local development of the multiplex offers better access to a diagnosis for respiratory infections, in addition to increasing research and improving technological capacity in order to obtain sanitary sovereignty in Ecuador.