FTA-LAMP based biosensor for a rapid in-field detection of Globodera pallida-the pale potato cyst nematode

Front Bioeng Biotechnol. 2024 Jan 18:12:1337879. doi: 10.3389/fbioe.2024.1337879. eCollection 2024.

Abstract

The combination of a sensitive and specific magnetoresistive sensing device with an easy DNA extraction method and a rapid isothermal amplification is presented here targeting the on-site detection of Globodera pallida, a potato endoparasitic nematode. FTA-cards were used for DNA extraction, LAMP was the method developed for DNA amplification and a nanoparticle functionalized magnetic-biosensor was used for the detection. The combinatorial effect of these three emerging technologies has the capacity to detect G. pallida with a detection limit of one juvenile, even when mixed with other related species. This combined system is far more interesting than what a single technology can provide. Magnetic biosensors can be combined with any DNA extraction protocol and LAMP forming a new solution to target G. pallida. The probe designed in this study consistently distinguished G. pallida (∆Vac binding/Vac sensor above 1%) from other cyst nematodes (∆Vac binding/Vac sensor below 1%). It was confirmed that DNA either extracted with FTA-cards or Lab extraction Kit was of enough quantity and quality to detect G. pallida whenever present (alone or in mixed samples), ensuring probe specificity and sensitivity. This work provides insights for a new strategy to construct advanced devices for pathogens in-field diagnostics. LAMP runs separately but can be easily integrated into a single device.

Keywords: FTA-card ®; LAMP; Lab-on-a-chip; PCN; magnetoresistive biochips.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by National Funds through FCT—Foundation for Science and Technology under the PhD fellowships BD 138724/2018 and COVID/BD/152764/2023, the project UIDB/05183/2020 (MED–Mediterranean Institute for Agriculture, Environment and Development, Institute for Advanced Studies and Research, Évora University) and the project UID/05367/2020 (PROGRAMATICO—Research Unit INESC MN, Microsystems and Nanotechnologies). Publication was supporteded by Bactometer, 2022-RIS_Innovation-068, funded through EIT Health.