American tegumentary leishmaniasis diagnosis using L. (V.) braziliensis fixed promastigotes: A comparative performance of serological tests and spontaneous cure identification

Abstract

Background: The present study aimed to demonstrate the applicability of a flow cytometry-based serology approach to identify spontaneous cure by the detection of immunoglobulin G, and also, the diagnosis and cure criterion by the IgG1 isotype in American Tegumentary Leishmaniasis-ATL caused by L. (V.) braziliensis. Also, a comparison between flow cytometry with the serological conventional technique was performed. Methods: Forty five individuals were included in study. They were assessed in two moments: First, 8 subjects spontaneously cured of ATL, 8 healthy individuals and 15 patients who had a positive diagnosis for ATL were selected before treatment to identify spontaneous cure by immunoglobulin G detection. Secondly, 14 patients who were positive for ATL were selected and had their blood collected before and 1, 2 and 5 years after treatment, respectively, for the diagnostic tests (ELISA and flow cytometry) and cure criterion evaluation using the IgG1 isotype. Results: The analysis of the mean percentage of positive fluorescent parasites (PPFP) along with the titration curves of IgG anti-fixed promastigotes of L.(V.)braziliensis, confirmed the applicability of this method for monitoring spontaneous cure in ATL with outstanding co-positivity (100%) and co-negativity (100%) performance indexes. Regarding the results of the comparison between flow cytometry and ELISA it was seen that there was a better accuracy of the first one in relation to the other. When IgG1 applicability was evaluated, it was observed that before treatment, 36.8% of the patients were negative; in patients 1 year post-Treatment, 82.3%; 2 years post-Treatment, 27.2% and in patients 5 years post-Treatment, 87.5%. The overall analysis of the results suggests that flow cytometry can be applied to ATL detection, and that the use of IgG1 isotype has possibilities to contribute as a more specific diagnostic method. Conclusions: Therefore, this area has great perspectives use for the diagnosis and cure criterion, and also it can be scaled up with the possibility to characterize the different clinical stages of the disease. Together, these findings demonstrate the applicability of a flow cytometry-based serology approach and opens up new avenues of research with this technique, such as the understanding the humoral response in ATL patients.