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Título: Standardization and validation of an indirect ELISA based on the extraction of proteins from Leptospira spp. as an antigen for the diagnosis of canine leptospirosis
Autor(es): Alves, Thaise
Fernandes, Bruno
Vieira, Vinícius
Pinheiro, Alexandre
Data do documento: 2023
Resumo: Leptospirosis is an important zoonosis. The World Health Organization recommends the use of microscopic soroagglutination in the diagnosis of the disease. However, this is a time-consuming technique that offers a risk of contamination to the performer and requires a trained professional. The indirect enzyme-linked immunosorbent assay (ELISA) test presents less risk, is faster, and has lower cost benefit for high sampling. In view of the advantages of using the test, and the importance of the development of techniques that facilitate and make the diagnosis more efficient, the objective of this study was to standardize an indirect ELISA technique using an immunogenic protein of Leptospira spp. for the diagnosis of canine leptospirosis. The standardized test obtained a cut off of 0.314, 76% of sensitivity, 34% of specificity, 53.52% of positive predictive value, 58.62 of negative predictive value, and 55% of accuracy. The protein extraction technique was efficient for antigen production, and the standardized test can be used in the diagnosis of canine leptospirosis.
Descrição: Leptospirosis is a zoonosis, and the diagnosis of infected animals is an important control of the disease. The etiologic agent is the bacterium Leptospira spp., which is part of the order Spirochaetales, family Leptospiraceae, and has 20 serogroups consisting of 250 serovars. The bacteria have cell wall and cytoplasmic membrane surrounded by the outer membrane containing lipoproteins (LigA, LigB, LenA, LenB, LcpA, Loa22, LipL32) and porins or transmembrane protein (OmpL1), and surface lipoproteins (FRAGA et al., 2016). The outer membrane surface lipoproteins present in pathogenic strains play an important role as an immunogen, being expressed both in in-vivo and in-vitro cultures, and such characteristics confer potential in the development of diagnostic tests for leptospirosis (JOSE et al., 2018). Notwithstanding, MONARIS et al. (2015), when studying outer membrane proteins of Leptospira interrogans, found that LigA can provide protective immunity. CASTIBLANCO-VALENCIA et al. (2016) found that LigA and LigB proteins are also related to the pathogen’s evasion mechanism from the immune system thanks to their ability to inactivate the complement system. FERNANDES (2012) highlights that the OmpL1 protein plays an important role in the hemorrhagic picture of leptospirosis infection, with competence in binding to plasminogen and fibronectin.
Palavras-chave: Leptospirosis
Indirect ELISA
Microscopic agglutination test
Diagnostic immunology
Citação: Alves, Thaise et al. Standardization and validation of an indirect ELISA based on the extraction of proteins from Leptospira spp. as an antigen for the diagnosis of canine leptospirosis. Arquivos do Instituto Biológico, 2023, v. 90, https://doi.org/10.1590/1808-1657000232022.
ISSN: 1808-1657
Número DOI: 10.1590/1808-1657000232022
Idioma: en_US
Editora: Instituto Biológico
Local da Publicação: São Paulo
Periodicidade: Fluxo contínuo
URI: http://repositoriobiologico.com.br//jspui/handle/123456789/1233
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