Prototyping and validation of an ELISA kit with recombinant Lb6H antigen for the serological diagnosis of American Cutaneous Leishmaniasis (ATL)

• Grant number: 21/12535-2
• Support Opportunities: Regular Research Grants
• Start date: April 01, 2022
• End date: March 31, 2025
• Field of knowledge: Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
• Principal Investigator: Maria Carmen Arroyo Sanchez
• Grantee: Maria Carmen Arroyo Sanchez
• Host Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Hiro Goto ; José Angelo Lauletta Lindoso

Abstract

The diagnosis of American Tegumentary Leishmaniasis (ATL) is an open question in our country, and the search for a solution is urgent. Laboratory tests are important in diagnosing ATL because the clinical manifestations, although often suggestive, are not pathognomonic, requiring a differential diagnosis. From laboratory tests, those aimed at finding parasites in samples from lesions are considered the gold standard, but the sensitivity is not high; molecular tests aimed at detecting Leishmania DNA still lack standardization among the various laboratories. In this scenario, there is the option of indirect laboratory tests that detect the presence of an immune response to Leishmania, serological examination, or delayed hypersensitivity test. However, the Montenegro test (delayed hypersensitivity), which was used in the clinical routine, is no longer performed due to discontinuity in the antigen production, causing problems in the diagnosis of ATL. In a recent study, our group evaluated ELISA with recombinant (r) Lb6H antigen derived from the Leishmania (Viannia) braziliensis sequence, which showed excellent performance, with a sensitivity of 100.0% and specificity of 98.5% for the diagnosis of this pathological condition. With these results and the lack of consensus regarding the ATL serology, we envision using rLb6H in the routine diagnosis. In this project, we will validate a serological test in ELISA format using the rLb6H antigen. We will then proceed with developing a prototype of an ELISA-rLb6H kit to complement the diagnosis of ATL. The importance of this project is evident, as it will come to fill when the ELISA test is validated and developed, the gap in the lack of a complementary diagnostic test for ATL. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

VALENCIA-PORTILLO, RUTH TAMARA; LINDOSO, JOSE ANGELO; CELESTE, BEATRIZ JULIETA; BITTENCOURT, AMANDA AZEVEDO; DE BRITO, MARIA EDILEUZA FELINTO; DUTHIE, MALCOLM SCOTT; GUDERIAN, JEFFERY; GUERRA, JORGE; OLIVEIRA, ANA LUCIA LYRIO; REED, STEVEN; et al. ELISA with recombinant antigen Lb6H validated for the diagnosis of American tegumentary leishmaniasis. PLoS One, v. 19, n. 6, p. 19-pg., 2024-06-05. (21/12535-2)

FUJIMORI, M.; VALENCIA-PORTILLO, R. T.; LINDOSO, J. A. L.; CELESTE, B. J.; DE ALMEIDA, R. P.; COSTA, C. H. N.. Recombinant protein KR95 as an alternative for serological diagnosis of human visceral leishmaniasis in the Americas (vol 18, e0282483, 2023). PLoS One, v. 18, n. 10, p. 1-pg., 2023-10-12. (21/12535-2, 17/03367-3)