Dados do Trabalho

Título

Evaluation of a hypothetical protein for the diagnosis of Tegumentary Leishmaniasis using patients urine samples

Introdução

Parasitological and molecular diagnoses of Tegumentary Leishmaniasis (TL) require invasive procedures, such as lesion biopsies, and are also limited in terms of test sensitivity since the presence of the parasite or genetic material in the tissue material is crucial. Although immunological tests are also used for the diagnosis, the essays present variable sensitivity and/or specificity, depending on the antigen used. Additionally, collecting blood to obtain serum for use can be unpleasant or inconvenient for those who undergo the procedure. In this sense, the use of urine for investigating antibodies for disease diagnosis could be considered as an alternative biological material, considering its easier collection when compared to blood, in addition to having simpler storage and handling.

Objetivo (s)

Evaluate the performance of a hypothetical protein from Leishmania mexicana, called AMT3, as an antigen in indirect enzyme-linked immunosorbent assays (ELISA) using urine and serum from patients with TL.

Material e Métodos

The hypothetical protein AMT3 was cloned, expressed, and purified. As a comparative antigen, Leishmania soluble antigenic extract (SLA) from L. amazonensis was used. For performing the indirect ELISA assays, these antigens were used against urine and serum samples from patients with TL: Cutaneous Leishmaniasis (CL; n=22) and Mucosal Leishmaniasis (ML; n=23). Samples from healthy individuals residing in endemic areas (n = 29) were used as a control.

Resultados e Conclusão

Sensitivity (Se), specificity (Sp), and Area Under the Curve (AUC) values of 95.56%, 100%, and 0.99, respectively, were observed for AMT3 using urine samples, while Se, Sp, and AUC values of 88.89%, 82.76%, and 0.89 using serum samples. Comparatively, Se, Sp, and AUC values of 81.82%, 100%, and 0.95 were observed for SLA, using urine samples, while for serum samples Se, Sp, and AUC values of 79.55%, 96.55%, and 0.86 were observed using serum samples. In conclusion, preliminary data from this study demonstrate that AMT3 is capable of detecting anti-leishmania antibodies in urine with higher performance when compared to the use of serum samples. Therefore, this urine-based ELISA using AMT3 could be considered a non-invasive method with potential application as an accessible platform for immunodiagnosis of tegumentary leishmaniasis.

Palavras-chave

Cutaneous Leishmaniasis; Diagnosis; Hypothetical Protein; Urine-based ELISA.

Agradecimentos

LIMT and LBAEL Teams; CAPES, CNPq, and FAPEMIG.