Specific and sensitive detection of Influenza A virus using a biotin-coated nanoparticle enhanced immunomagnetic assay
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Specific and sensitive detection of Influenza A virus using a biotin-coated nanoparticle enhanced immunomagnetic assay Carole Farre 1 & Sara Viezzi 1,2 & Alice Wright 1 & Perrine Robin 1 & Nathalie Lejal 3 & Marisa Manzano 2 & Jasmina Vidic 4 & Carole Chaix 1 Received: 1 October 2020 / Revised: 4 November 2020 / Accepted: 17 November 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract This study reports the development of a sensitive magnetic bead-based enzyme-linked immunoassay (MELISA) for the panreactive detection of the Influenza A virus. The assay combines immunomagnetic beads and biotin-nanoparticle-based detection to quantify a highly conserved viral nucleoprotein in virus lysates. At the capture step, monoclonal antibody-coated magnetic microbeads were used to bind and concentrate the nucleoprotein in samples. The colorimetric detection signal was amplified using biotinylated silica nanoparticles (NP). These nanoparticles were functionalized on the surface with short DNA spacers bearing biotin groups by an automated supported synthesis method performed on nano-on-micro assemblies with a DNA/RNA synthesizer. A biotin-nanoparticle and immunomagnetic bead-based assay was developed. We succeeded in detecting Influenza A viruses directly in the lysis buffer supplemented with 10% saliva to simulate the clinical context. The biotin-nanoparticle amplification step enabled detection limits as low as 3 × 103 PFU mL−1 and 4 × 104 PFU mL−1 to be achieved for the H1N1 and H3N2 strains respectively. In contrast, a classical ELISA test based on the same antibody sandwich showed detection limit of 1.2 × 107 PFU mL−1 for H1N1. The new enhanced MELISA proved to be specific, as no cross-reactivity was found with a porcine respiratory virus (PRRSV). Keywords Immunoassay . Influenza A virus . Magnetic bead . Biotin-nanoparticle . Nucleoprotein . Diagnostics
Introduction Influenza viruses cause worldwide outbreaks and pandemics in humans and animals with high morbidity and mortality [1]. Of the Influenza A, B, C and D viruses, Influenza A viruses mutate the most rapidly leading to the continuous apparition of new strains. The high antigenic flexibility and hence greater Published in the topical collection celebrating ABCs 20th Anniversary. * Carole Chaix [email protected] 1
Université de Lyon, CNRS, Université Claude Bernard Lyon 1, Institut des Sciences Analytiques, UMR 5280, 5 rue de la Doua, 69100 Villeurbanne, France
2
Dipartimento di Scienze Agro-Alimentari, Ambientali e Animali, Università degli Studi di Udine, via Sondrio 2/A, 33100 Udine, Italy
3
Université Paris-Saclay, UR892, INRAE, 78350 Jouy-en-Josas, France
4
Université Paris-Saclay, INRAE, AgroParisTech, Micalis Institute, 78350 Jouy-en-Josas, France
virulence increase the severity of illnesses and death rates in the event of a pandemic [2]. The seasonal subtypes of circulating Influenza A viruses (H1N1 and H3N2) cause 3–5 million human severe infections and between 290,000 and 650,000 fatal case
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