Design and validation of a semi-quantitative microneutralization assay for human Metapneumovirus A1 and B1 subtypes
Since 2001, human Metapneumovirus has been a significant cause of human respiratory disease worldwide, and no vaccine or preventive treatment is currently available. The ELISA-based live virus microneutralization assay is a method to detect neutralizing antibodies against a target pathogen. The aim of this study was to demonstrate the suitability of this approach to quantifying neutralizing antibodies against A1 and B1 virus subtypes in human serum samples. To standardize and validate this microneutralization assay, we carried out analytical procedures according to the International Council of Harmonization guidelines; these procedures are described in detail. In addition, we compared the validated method with the indirect ELISA, and confirmed that the ELISA-based microneutralization assay provides reliable, accurate and reproducible results. The use of this high-throughput method for large-scale serological studies could effectively support the evaluation of the immunogenicity of new vaccines, thereby improving therapeutical strategies against human Metapneumovirus.
Authors
- Angela Stufano (ORCID: https://orcid.org/0000-0002-9101-7742)
- Mariangela Pizzo (ORCID: https://orcid.org/0000-0001-7790-2828)
- Simona De Grazia (ORCID: https://orcid.org/0000-0001-7748-0880)
- Francesca Dapporto (ORCID: https://orcid.org/0000-0001-7897-4452)
- Maria Alfreda Stincarelli (ORCID: https://orcid.org/0000-0003-1046-4565)
- Carolina Bonifazi
- Claudia Maria Trombetta (ORCID: https://orcid.org/0000-0001-7229-9919)
- Roberta Antonelli (ORCID: https://orcid.org/0000-0001-6884-7847)
- Pietro Piu (ORCID: https://orcid.org/0000-0002-7504-1822)
- Ginevra Giglioli
- Alessandro Manenti (ORCID: https://orcid.org/0000-0002-4027-7296)
- Giulia Riolo (ORCID: https://orcid.org/0000-0003-2888-5755)
- Emanuele Montomoli (ORCID: https://orcid.org/0000-0001-7595-4974)
- Piero Lovreglio (ORCID: https://orcid.org/0000-0002-1609-9397)
- Giulia Roscia (ORCID: https://orcid.org/0009-0001-6999-0175)
- Noemi Guerrini (ORCID: https://orcid.org/0009-0001-8998-9446)
- Valentina Biagini
Institutions
- University of Siena (IT)
- VisMederi Research (IT)
- University of Bari Aldo Moro (IT)
- University of Palermo (IT)
Publication Details
- Journal
- Scientific Reports
- Published
- 2025-04-04
- DOI
- https://doi.org/10.1038/s41598-025-96567-6
- Citations
- 2
- Primary Topic
- Respiratory viral infections research
- Type
- article
- Field-Weighted Citation Impact
- 2.71
Funders
- Erasmus Medisch Centrum