Fluorescence quantification of bacterial endospores using aptazyme pairs and molecular beacons

IGF 21043 N

The aim of the research project is to develop a method for the simultaneous quantification of endospores (total number of spores) and endospores of hygiene-relevant bacteria (e.g. C. difficile, B. cereus): This can be realized by developing pairs of aptazymes and molecular beacons specially designed as nucleozyme substrates, whereby the aptazymes of a pair cleave the same substrate.

The first aptazyme of a pair is activated upon binding of a primary target molecule (concentration proportional to the number of spores) and cleaves the Molecular Beacon, releasing a fluorophore and a quencher. The released quencher serves as induced target molecule for the second aptazyme.

Thus, the number of induced target molecules doubles with each substrate cleavage, which leads to an exponential increase in the number of induced target molecules and released fluorophores with increasing incubation time (exponential signal increase).

The project results will provide the cleaning service providers and the textile service providers with a method that will enable them for the first time to evaluate the spore contamination of surfaces, textiles and process water within the scope of in-house self-monitoring and thus to monitor the success of decontamination measures they have taken.

If endospores (e.g. C. difficile, B. cereus) are detected, suitable measures for spore decontamination can be taken immediately and hygiene safety can be further optimized.

The IGF-pro­ject 21043 N of the re­search as­so­ci­a­tion Europäische Forschungs­ge­mein­schaft Reini­gungs- und Hy­gien­etech­nolo­gie e.V., Cam­pus Ficht­en­hain 11, 47807 Krefeld, is sup­port­ed via the AiF with­in the fund­ing pro­gram „In­dus­trielle Gemein­schafts­forschung und -en­twick­lung (IGF)“ by the Fed­er­al Min­istry of Eco­nom­ic Af­fairs and En­er­gy (BMWi) due to a de­ci­sion of the Ger­man Par­lia­ment.