Qualitative Real‐Time Disinfection Monitoring through Lipid Nanoparticle‐Separated Fluorophore‐Quencher Pairs

Abstract Clean, disinfected surfaces and medical instruments are critical to maintaining a hygienic environment, especially in healthcare settings. Current methods for disinfection validation and training require either a long evaluation time or do not distinguish between physical (dilution) and chemical (disintegration) disinfection procedures. However, to achieve effective disinfection, both effects, dilution and disintegration, are required for many commonly used disinfectants (e.g., alcohol, sodium hypochlorite, quaternary ammonium compounds). In this study, a method is established for the real‐time monitoring of surface disinfection using fluorescence‐labeled DNA and lipid nanoparticles (LNP) encapsulating such DNA. It is shown that the spatial separation of quencher‐modified DNA and fluorophore‐modified complementary DNA by LNPs can be disrupted by ethanolic disinfectants, facilitating the disintegration of LNPs. The resulting quenching of fluorescence can immediately be detected using a manual setup comprising a hand‐held laser, a color filter, and a smartphone camera. To demonstrate a potential application of this novel disinfection detection technology, disinfection of a commonly used medical instrument, a scalpel, is validated using the qualitative change in fluorescence upon disintegration of LNPs, enabling distinction between physical dilution and chemical disintegration. Therefore, LNPs spatially separating quencher and fluorophore offer real‐time, qualitative monitoring of surface disinfection.