Electrochemical molecular beacon DNA biosensor for the detection and discrimination of the DF508 cystic fibrosis mutation

Cystic fibrosis is one of the most common genetically inherited diseases in Northern Europe, consisting of a defect of chloride transport in the epithelium, with the DF508 mutation being the most common mutation associated with the disease. In this work the design and characterisation of a reagent-less electrochemical genosensor, based on the use of an electrochemical molecular beacon targeting the DF508 mutation is presented. Different aspects of the sensing platform including molecular beacon design and surface chemistry of the sensor surface were evaluated. Operational parameters such as assay buffer and assay time were also optimised. Using the optimised molecular beacon designs a clear differentiation between the targeted sequence (i.e. mutant) and potential interferent (i.e. wild type) was demonstrated, with a total required assay time of 20 min. The major advantage of the proposed reagent-less sensing platform is the fact that this only required, as intervention of the end-user, the addition of the sample.

► The design of an electrochemical molecular beacon for DF508 mutation has been defined.
► Optimisation and stability of on-surface immobilisation of the MB were evaluated.
► Optimal assay conditions, time and buffer, have been identified.
► Discrimination between mutant amplicon and wild type amplicon demonstrated.