CRISPR-Cas13a-Based Assay for Accurate Detection of OXA-48 and GES Carbapenemases
Concha Ortiz-Cartagena, Daniel Pablo‐Marcos, Laura Fernández-García, Lucía Blasco, Olga Pacios, Inés Bleriot, María Siller, María López, Javier Fernández, Belén Aracil, Pablo A. Fraile-Ribot, Sergio García‐Fernández, Felipe Fernández-Cuenca, Marta Hernández-García, Rafael Cantón, Jorge Calvo, María Tomás
Abstract
Carbapenems are one of the last-resort antibiotics for defense against multidrug-resistant pathogens. Multiple nucleic acid amplification methods, including multiplex PCR, multiplex loop-mediated isothermal amplification (LAMP) and multiplex RPAs, can achieve rapid, accurate, and simultaneous detection of several resistance genes to carbapenems in a single reaction. However, these assays need thermal cycling steps and specialized instruments, giving them limited application in the field. In this work, we adapted with high specificity and sensitivity values, a new LAMP CRISPR-Cas13a-based assay for the detection of OXA-48 and GES carbapenemases in clinical samples without the need for RNA extraction.