Dados do Trabalho

Título

Development and evaluation of a rapid test based on the recombinant structural protein VP1-2A for detecting acute infections of hepatitis A virus.

Introdução

In recent years, the epidemiological profile of hepatitis A in Brazil has changed, with endemicity shifting from high to moderate. As a result, epidemic outbreaks have been reported, particularly targeting young people and adults who are at a higher risk of morbidity and mortality from HAV. Given this scenario, rapid tests (RT) that can provide an immediate result without requiring specialized training or equipment, could be a powerful tool for serological tracking of acute hepatitis cases. This technique could be especially valuable in remote areas lacking laboratory facilities, improving diagnosis accessibility.

Objetivo (s)

The goal of this research was to create a rapid test (RT) using a lateral flow immunochromatography platform to detect antibodies against acute hepatitis A infection (anti-HAV IgM).

Material e Métodos

To achieve this, a recombinant VP1-2A protein was expressed in Eschericha coli, purified using immobilized metal ion affinity chromatography (IMAC), and a high level of the protein was obtained. The recombinant VP1-2A protein underwent antigenic evaluation using anti-HAV control samples (positive and negative) through Multi-Antigen Print Immunoassay (MAPIA) and liquid microarray. This study was approved at Ethics Committee of the Oswaldo Cruz Foundation guidelines (No. 1.610.747).

Resultados e Conclusão

The recombinant protein exhibited anti-HAV IgM binding at different concentrations (40, 50, 60, 80 and 100 ng/mm/band), demonstrating its antigenicity. No cross-reaction was observed in negative anti-HAV samples. These findings indicate that the recombinant protein can be used in rapid test development for the detection of anti-HAV IgM antibodies. The standardization of rapid tests started with the evaluation of different parameters such as running buffer, antigen concentration, and porosity of the nitrocellulose membrane. To validate the performance of the rapid test, a panel of serum samples characterized for anti-HAV by Chemiluminescense immunoassay (CLIA) was used (n=89). The developed rapid test showed a high agreement with CLIA results (80%). The rapid test has the characteristics of high accuracy and good sensitivity and specificity (70% and 86%, respectively), making it suitable for the rapid screening of acute hepatitis A. This result shows that our RT can be used as an alternative for diagnosing hepatitis A in various settings, including remote and difficult-to-access locations

Palavras Chave

Hepatitis A; Poin-of-care tests; diagnosis; recombinant protein.