Magnetic Field-Enhanced Agglutination Readout Combined With Isothermal Reverse Transcription Recombinase Polymerase Amplification for Rapid and Sensitive Molecular Detection of Dengue Virus.

Among the numerous molecular diagnostic methods, isothermal reverse transcription recombinase polymerase amplification (RT-RPA) is a simple method that has high sensitivity and avoids the use of expensive instruments. However, detection of amplified genomes often requires a fluorescence readout on costly readers or migration on a lateral flow strip with a subjective visual reading. Aiming to establish a new approach to rapidly and sensitively detect viruses, we combined RT-RPA with a magnetic field-enhanced agglutination (MFEA) assay and assessed the ability of this method to detect the dengue virus (DENV). Magnetization cycles accelerated the capture of amplified DENV genomes between functionalized magnetic nanoparticles by a fast chaining process to less than 5 min; the agglutination was quantified by simple turbidimetry. A total of 37 DENV RNA

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Copyright © 2022 Leon, Pinchon, Mayran, Daynès, Morvan, Molès, Cantaloube and Fournier-Wirth.

Déclaration de conflit d'intérêts

AD is an employee of HORIBA Medical. FL, EP, CM, AD, J-PM, J-FC, and CF-W are coinventors of patent PCT/FR2021/000022 (Detection of infectious agents based on recombinase polymerase amplification combined with magnetic field-enhanced agglutination). The remaining author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The handling Editor declared a past co-authorship with one of the authors (CF). The authors declare that this study received funding from HORIBA Medical, Etablissement Français du Sang (EFS), and by the University of Montpellier and its I-SITE MUSE Program (ArboMag project) funded by the french government. HORIBA Medical funder had the following involvement in the study: i) AD is a company employee with expertise on magnetic agglutination, ii) the prototype platform for magnetic field-enhanced agglutination is provided by the company.