Affiliation:
1. Laboratório de Virologia Molecular Instituto Carlos Chagas (ICC/Fiocruz) Curitiba Paraná Brasil
2. Complexo Hospital de Clínicas Universidade Federal Do Paraná Curitiba Paraná Brasil
3. Laboratório de Patologia Experimental Pontifícia Universidade Católica do Paraná Curitiba Paraná Brasil
4. Laboratório de Biologia Estrutural e Engenharia de Proteínas Instituto Carlos Chagas (ICC/Fiocruz) Curitiba Paraná Brasil
Abstract
AbstractSince its emergence in late 2019, coronavirus disease 2019 (COVID‐19) has caused millions of deaths and socioeconomic losses. Although vaccination significantly reduced disease mortality, it has been shown that protection wanes over time, and that severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) variants of concern (VOCs) may escape vaccine‐derived immunity. Therefore, serological studies are necessary to assess protection in the population and guide vaccine regimens. A common measure of protective immunity is the presence of neutralizing antibodies (nAbs). However, the gold standard for measuring nAbs (plaque reduction neutralization test, or PRNT) is laborious and time‐consuming, limiting its large‐scale applicability. We developed a high‐throughput fluorescence reduction neutralization assay (FRNA) to detect SARS‐CoV‐2 nAbs. Because the assay relies on immunostaining, we developed and characterized monoclonal antibodies (mAbs) to lower costs and reduce the assay's vulnerability to reagent shortages. Using samples of individuals vaccinated with COVID‐19 and unvaccinated/pre‐pandemic samples, we showed that FRNA results using commercial and in‐house mAbs strongly correlated with those of the PRNT method while providing results in 70% less time. In addition to providing a fast, reliable, and high‐throughput alternative for measuring nAbs, the FRNA can be easily customized to assess SARS‐CoV‐2 VOCs. Additionally, the mAb we produced was able to detect SARS‐CoV‐2 in pulmonary tissues by immunohistochemistry assays.
Funder
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Subject
Infectious Diseases,Virology
Reference33 articles.
1. A Novel Coronavirus from Patients with Pneumonia in China, 2019
2. COVID ‐ Coronavirus Statistics ‐ Worldometer. Accessed February 13 2023.https://www.worldometers.info/coronavirus/#countries
3. Clinical features of patients infected with 2019 novel coronavirus in Wuhan, China
4. Centers for Disease Control and Prevention.People with Certain Medical Conditions | CDC. Accessed February 13 2023.https://www.cdc.gov/coronavirus/2019-ncov/need-extra-precautions/people-with-medical-conditions.html
5. Coronavirus disease (COVID‐19): comprehensive review of clinical presentation;Mehta OP;Front Public Heal,2021