Author | Simões, Marisol | |
Author | Silva, Stephanie Almeida da | |
Author | Lúcio, Kelly Araújo | |
Author | Vieira, Renan de Oliveira | |
Author | Schwarcz, Waleska Dias | |
Author | Lima, Sheila Maria Barbosa de | |
Author | Camacho, Luiz Antonio Bastos | |
Access date | 2024-04-29T14:45:31Z | |
Available date | 2024-04-29T14:45:31Z | |
Document date | 2023 | |
Citation | SIMÕES, Marisol et al. Standardization, validation, and comparative evaluation of a faster and high-performance test for quantification of yellow fever neutralizing antibodies. Journal of Immunological Methods, v. 522, p. 1-12, Nov. 2023. | en_US |
ISSN | 0022-1759 | en_US |
URI | https://www.arca.fiocruz.br/handle/icict/63715 | |
Language | eng | en_US |
Publisher | Elsevier | en_US |
Rights | open access | en_US |
Title | Standardization, validation, and comparative evaluation of a faster and high-performance test for quantification of yellow fever neutralizing antibodies | en_US |
Type | Article | en_US |
DOI | 10.1016/j.jim.2023.113568 | |
Abstract | Although it is considered the reference for quantification of neutralizing antibodies, classical method of the plaque reduction neutralization test (PRNT) is labor intensive, requires specific equipment and inputs, besides a long time for its finalization, even in the micro-PRNT version (in 96-well plates). It has a higher sample throughput, however the smaller wells make the reading of plaques more difficult. With an immunoenzymatic revelation step and a semi-automated reading, the μ μ FRN-HRP (micro Focus Reduction Neutralization - Horseradish Peroxidase) is a faster and more efficient test for the quantification of YF neutralizing antibodies. This study aimed to standardize, validate, and compare it with the reference method in 6-well plates (PRNT). Once the execution protocol was standardized, precision, accuracy, selectivity, and robustness were evaluated to validate the FRN-HRP. In addition, 200 sera of vaccinees were processed by the μ FRN-HRP and by the micro-PRNT to compare with the reference test, estimating agreement by Intraclass Correlation Coefficient (ICC). The standardization and validation of the μ FRN-HRP was carried out successfully. Weak to moderate agreement was observed between μ FRN-HRP and PRNT for titers in reciprocal dilution, while the same comparison between the classical tests resulted in a better ICC. However, titers in milli-international units obtained by μ μ FRN-HRP showed a substantial agreement with PRNT, while the agreement between micro-PRNT and PRNT was inferior. Therefore, FRN-HRP can be used in the confirmation of natural YF infection and immune response to vaccination, replacing the micro-PRNT, gaining agility, while preserving the specificity of the result. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto de Tecnologia em Imunobiológicos (Bio-Manguinhos). Laboratório de Tecnologia Virológica. Rio de Janeiro, RJ, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Escola Nacional de Saúde Pública Sergio Arouca. Rio de Janeiro, RJ, Brasil. | en_US |
Subject | FRNT | en_US |
Subject | PRNT | en_US |
Subject | Neutralizing antibodies | en_US |
Subject | Yellow fever virus | en_US |
Subject | Standardization | en_US |
Subject | Validation | en_US |