Multiplex diagnostics: fast, broad and reliable

“Multiplex luminescent diagnostics for infectious diseases”

Nucleic acid detection methods based on CRISPR and isothermal amplification techniques offer promising opportunities for point-of-care diagnostics of infectious diseases. We have LUNAS (Luminescent Nucleic Acid Sensor) platform was developed, in which target dsDNA is sequence-specifically detected by a pair of dCas9-based probes that enable split luciferase complementation. LUNAS is easily combined with recombinase polymerase amplification (RPA) and can be interrogated with a simple digital camera.

To enable multiplex detection, we developed LUNAS color variants that can detect and distinguish two pathogens in a single reaction. For demonstration purposes, we designed a duplex assay for N. gonorrhoeae and C. trachomatis, two sexually transmitted bacterial infections with similar clinical symptoms. We validated the accuracy of this test with patient samples, demonstrating the potential of this method for rapid and reliable multiplex diagnostics of infectious diseases.

Speaker: Yosta de Stigter, TU Eindhoven