| Autor: |
Franco-Urquijo PA; Laboratorio de Terapia Génica, Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Mexico City 07360, Mexico., Sierra-Martínez M; Unidad de Investigación en Salud, Hospital Regional de Alta Especialidad de Ixtapaluca, Ixtapaluca 56530, Mexico., Jarquín-Martínez M; Unidad de Investigación en Salud, Hospital Regional de Alta Especialidad de Ixtapaluca, Ixtapaluca 56530, Mexico., Martínez-Roque MA; Laboratorio de Terapia Génica, Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Mexico City 07360, Mexico., García-Velásquez VM; Laboratorio de Terapia Génica, Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Mexico City 07360, Mexico., Acosta-Altamirano G; Hospital Regional de Alta Especialidad de Ixtapaluca, Ixtapaluca 56530, Mexico., Ruiz-Pérez NJ; Independent Researcher, Mexico City 07800, Mexico., Toscano-Garibay JD; Unidad de Desarrollo en Soluciones Diagnósticas, Hospital Regional de Alta Especialidad de Ixtapaluca, Ixtapaluca 56530, Mexico., Alvarez-Salas LM; Laboratorio de Terapia Génica, Departamento de Genética y Biología Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Mexico City 07360, Mexico. |
| Abstrakt: |
The COVID-19 pandemic has been a main concern over the last two years and has become one of the most important crises in the history of human health. Today, there is still a need for affordable and reliable diagnostic tests for massive disease monitoring. Previously, a set of highly specific DNA-aptamers (C7/C9) binding to the SARS-CoV-2 Spike (S) protein were isolated but its performance in clinical samples remained to be tested. Here, 242 samples were collected through three different methods and subjected to florescence-linked aptamer assays (FLAA) based on C7/C9 aptamers through two readout protocols. Then, a step-by-step statistical approach which included agreement tests, proportion comparisons and binomial and multinomial logistic regressions was used to predict optimal conditions for the novel C7/C9 FLAA test. RTqPCR threshold cycles, symptoms onset and processing time were influential factors on FLAA test results. Naturally occurring mutations on S were also detected and analyzed. Aminoacidic substitutions D614G and T732A appeared relevant for aptamer recognition although further studies are necessary. The methodology presented here is the first step to determine the performance and diagnosis across a range of clinical contexts and it might serve as a base for a complete analysis applicable to other designs of new diagnostic tests. |
