Convergence amplification and fluoroswitch detection for the determination of hygiene-relevant ESKAPE pathogens

IGF 21186 N

The aim is a cultivation independent simultaneous detection of all six nosocomial ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.) to control the hygiene status of disinfected surfaces in medical facilities.

The simultaneous detection of individual cells is based on an isothermal convergence amplification of specific DNA target sequences, which produces a uniform amplification product (amplicon) as a sum parameter. The amplicon is detected by an immobilized fluoroswitch, in which the amplicon binds to a complementary DNA immobilized on the surface of gold nanoparticles and carries a quantum dot on its free end.

Due to the conformational proximity between the quantum dot and the gold particles, the fluorescence of the dot is suppressed (switch off). When the amplicon binds, the forced conformation of the DNA is cancelled; the DNA stretches and the quantum dot shows a strong fluorescence due to the increased distance to the gold surface (switch on).

The Fluoroswitch is regenerable and can be used for numerous detection reactions. The detection allows the detection of single cells of ESKAPE pathogens within 45 minutes.

The cleaning and hygiene service providers, which are increasingly entrusted with the disinfection of surfaces in medical facilities, are provided with an in-house and cost-effective rapid detection system for independent control of the hygiene status of disinfected surfaces, which results in economic advantages.

The certificate improves hygiene in medical facilities which are required to disinfect surfaces in accordance with § 23 of the German Infection Protection Act (IfSG) and reduces the risk of nosocomial infections.

The IGF-pro­ject 21186 N of the re­search as­so­ci­a­tion Europäische Forschungs­ge­mein­schaft Reini­gungs- und Hy­gien­etech­nolo­gie e.V., Cam­pus Ficht­en­hain 11, 47807 Krefeld, was sup­port­ed via the AiF with­in the fund­ing pro­gram „In­dus­trielle Gemein­schafts­forschung und -en­twick­lung (IGF)“ by the Fed­er­al Min­istry of Eco­nom­ic Af­fairs and Cli­mate Ac­tion due to a de­ci­sion of the Ger­man Par­lia­ment.