Fluorescence quantification of bacterial endospores using aptazyme pairs and molecular beacons

IGF 21043 N

The aim of the research project was to develop a method for the simultaneous quantification of endospores (total spore number) and endospores of hygiene-relevant bacteria (e.g. C. difficile, B. cereus).

This can be realized by developing aptazyme pairs 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 spore number) and cleaves the molecular beacon, releasing a fluorophore and a quencher.

The released quencher serves as an induced target molecule for the second aptazyme. Thus, the number of induced target molecules doubles with each substrate cleavage, leading to the exponential increase in the number of induced target molecules and released fluorophores with increasing incubation time (exponential signal increase).

In the future, the project results will provide cleaning service providers and textile service providers with a method which, for the first time, will enable them to evaluate the spore load of surfaces, textiles and process waters within the framework of in-house self-monitoring and thus to monitor the success of decontamination measures they have undertaken.

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

The research report is available on request from FRT.

The IGF-pro­ject 21043 N of the re­search as­so­ci­a­tion Europäische Forschungs­ge­mein­schaft Reini­gungs- und Hy­gien­etech­nolo­gie e.V., Cam­pus Ficht­en­hain 11, 47807 Krefeld, was sup­port­ed 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.