PNA-functionalized magnetic microbeads as substrates for enzyme-labelled voltammetric genoassay for DNA sensing applied to identi fi cation of GMO in food

A novel enzyme-labelled voltammetric magnetogenoassay for DNA sensing based on the use of carboxyl surface coated magnetic microbeads functionalized with PNA probes and subsequent read-out on screen-printed electrode (SPE) substrates was developed. The assay was validated for determination of non-amplified genomic DNA from genetically modified Roundup Ready soy. Outstanding performance with respect to other genoassays requiring preliminary amplification of target DNA via PCR was demonstrated. The analytical performance was also improved compared to previous methods based on the immobilization of the same PNA probes on SPE substrates, since the method was found capable of achieving LOD and LOQ of 415 fM and 995 fM, respectively. The ability of the magnetogenoassay to detect the presence of Roundup Ready soy DNA sequence was tested on genomic DNA extract from European Reference Material soy flours, demonstrating the capability of the method to match the European Union regulation for labelling of food containing a percentage of GM products greater than 0,9%. ? 2021 Elsevier B.V. All rights reserved.

Automated summary

A novel enzyme-labelled voltammetric magnetogenoassay for DNA sensing using carboxyl surface coated magnetic microbeads functionalized with PNA probes has been developed, demonstrating outstanding performance without the need for PCR amplification of target DNA. The method showed improved analytical performance compared to previous methods based on the immobilization of the same PNA probes on SPE substrates, with LOD and LOQ levels achieved at 415 fM and 995 fM, respectively.