Implementación de un diagnóstico tipo ELISA para detección de anticuerpos contra la espícula de SARS-CoV-2

Abstract

The emergence of COVID-19 led to a shortage of reagents and tests due to the demand for diagnostics against this disease. On the other hand, along with vaccination, there is a need to follow up the immunity of the population. For this reason, an ELISA type diagnosis is sought to be implemented locally to detect antibodies against the Spike of this virus. This proposal fulfills the objective of Good Health and Well Being of the UN and will allow to be prepared for the need of booster doses and diagnosis of future variants. A region of the spike was selected and cloned into a vector suitable for mammalian cells. It was transfected into HEK293 cells. Two Western Blots were performed, testing an AntiFlag and a positive serum against the virus. The plasmid construct was successfully obtained and verified by sequencing. The protein was detected only by the virus-positive serum; however, its size was smaller than expected. For the design of the ELISA assay, the variables to be considered were established, and a base assay was proposed. The obtaining of the genetic construct and the design of the ELISA represent a great achievement for the implementation of this immunodiagnostic locally, which will lead Ecuador to the sanitary sovereignty the country needs