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Article

Characterization of Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Antibodies against Salmonella Typhimurium and Salmonella Enteritidis O-Antigens in Human Sera

by
Maria Grazia Aruta
1,
Elisa Lari
2,
Daniele De Simone
1,
Bianca Semplici
2,
Claudia Semplici
2,
Helen Dale
3,4,5,
Esmelda Chirwa
3,4,5,
Innocent Kadwala
3,4,
Maurice Mbewe
3,4,
Happy Banda
3,4,
Miren Iturriza-Gomara
1,
Melita Gordon
3,5,
Tonney Nyirenda
4,
Pietro Piu
2,
Mariagrazia Pizza
1,6,
Francesco Berlanda Scorza
1,
Silvia Grappi
2,
Rocío Canals
1,
Omar Rossi
1,* and
on behalf of the Vacc-iNTS Consortium Collaborators
†
1
GSK Vaccines Institute for Global Health (GVGH) S.r.l., 53100 Siena, Italy
2
VisMederi S.r.l., 53100 Siena, Italy
3
Malawi Liverpool Wellcome Trust Programme, Blantyre 30096, Malawi
4
Pathology Department, Kamuzu University of Health Sciences, Blantyre 312225, Malawi
5
Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L69 3BX, UK
6
Imperial College South Kensington Campus, London SW7 2AZ, UK
*
Author to whom correspondence should be addressed.
†
Collaborators/Membership of the Group/Team Name is provided in the Acknowledgments.
BioTech 2023, 12(3), 54; https://doi.org/10.3390/biotech12030054
Submission received: 18 July 2023 / Revised: 4 August 2023 / Accepted: 9 August 2023 / Published: 11 August 2023
(This article belongs to the Section Medical Biotechnology)
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Abstract

Nontyphoidal Salmonella (NTS) is a leading cause of morbidity and mortality caused by enteric pathogens worldwide in both children and adults, and vaccines are not yet available. The measurement of antigen-specific antibodies in the sera of vaccinated or convalescent individuals is crucial to understand the incidence of disease and the immunogenicity of vaccine candidates. A solid and standardized assay used to determine the level of specific anti-antigens IgG is therefore of paramount importance. In this work, we presented the characterization of a customized enzyme-linked immunosorbent assay (ELISA) with continuous readouts and a standardized definition of EU/mL. We assessed various performance parameters: standard curve accuracy, dilutional linearity, intermediate precision, specificity, limits of blanks, and quantification. The simplicity of the assay, its high sensitivity and specificity coupled with its low cost and the use of basic consumables and instruments without the need of high automation makes it suitable for transfer and application to different laboratories, including resource-limiting settings where the disease is endemic. This ELISA is, therefore, fit for purpose to be used for quantification of antibodies against Salmonella Typhimurium and Salmonella Enteritidis O-antigens in human samples, both for vaccine clinical trials and large sero-epidemiological studies.
Keywords: Generalized Modules for Membrane Antigens (GMMA); Enzyme-Linked Immunosorbent Assay (ELISA); non-typhoidal Salmonella; human sera; antibodies; vaccine; iNTS Generalized Modules for Membrane Antigens (GMMA); Enzyme-Linked Immunosorbent Assay (ELISA); non-typhoidal Salmonella; human sera; antibodies; vaccine; iNTS

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MDPI and ACS Style

Aruta, M.G.; Lari, E.; De Simone, D.; Semplici, B.; Semplici, C.; Dale, H.; Chirwa, E.; Kadwala, I.; Mbewe, M.; Banda, H.; et al. Characterization of Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Antibodies against Salmonella Typhimurium and Salmonella Enteritidis O-Antigens in Human Sera. BioTech 2023, 12, 54. https://doi.org/10.3390/biotech12030054

AMA Style

Aruta MG, Lari E, De Simone D, Semplici B, Semplici C, Dale H, Chirwa E, Kadwala I, Mbewe M, Banda H, et al. Characterization of Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Antibodies against Salmonella Typhimurium and Salmonella Enteritidis O-Antigens in Human Sera. BioTech. 2023; 12(3):54. https://doi.org/10.3390/biotech12030054

Chicago/Turabian Style

Aruta, Maria Grazia, Elisa Lari, Daniele De Simone, Bianca Semplici, Claudia Semplici, Helen Dale, Esmelda Chirwa, Innocent Kadwala, Maurice Mbewe, Happy Banda, and et al. 2023. "Characterization of Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Antibodies against Salmonella Typhimurium and Salmonella Enteritidis O-Antigens in Human Sera" BioTech 12, no. 3: 54. https://doi.org/10.3390/biotech12030054

APA Style

Aruta, M. G., Lari, E., De Simone, D., Semplici, B., Semplici, C., Dale, H., Chirwa, E., Kadwala, I., Mbewe, M., Banda, H., Iturriza-Gomara, M., Gordon, M., Nyirenda, T., Piu, P., Pizza, M., Berlanda Scorza, F., Grappi, S., Canals, R., Rossi, O., & on behalf of the Vacc-iNTS Consortium Collaborators. (2023). Characterization of Enzyme-Linked Immunosorbent Assay (ELISA) for Quantification of Antibodies against Salmonella Typhimurium and Salmonella Enteritidis O-Antigens in Human Sera. BioTech, 12(3), 54. https://doi.org/10.3390/biotech12030054

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