Multiplex diagnostics: fast, broad and reliable

Multiplex diagnostics enables the simultaneous detection of multiple pathogens in a single analysis. This approach saves time, reduces costs and increases efficiency in both clinical and research settings. During this seminar, you will discover how innovative techniques such as CRISPR-based detection and isothermal amplification are used for rapid and sensitive point-of-care diagnostics. We will explain the LUNAS platform: a luminescent nucleic acid test that can distinguish multiple pathogens in a single reaction thanks to color variants. This session offers an up-to-date and practice-oriented overview of the possibilities and challenges of multiplex diagnostics for infectious diseases, with attention to lab applications, data acquisition and implementation in healthcare.

“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

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