Validation of a Real-Time PCR for the Diagnosis of Leishmania Species Using the Hsp20 Gene

Maldonado-Aroni M.; Rojas-Palomino N.; Sandoval-Juarez A.; Galarza-Pérez M.; Alarcón-Guerrero J.; Guevara-Montero R.; Cárdenas-López V. · Tropical Medicine and Infectious Disease (2025) · DOI: 10.3390/tropicalmed10050121

Resumen

Leishmaniasis is a complex neglected tropical disease that impacts public health, particularly in resource-limited populations where access to accurate and timely diagnosis is often limited. Current diagnostic methods, primarily relying on microscopy, suffer from low sensitivity and specificity, hindering effective case management and disease control. The objective of this study was to validate a novel real-time PCR assay targeting the conserved Hsp20 gene for the detection of Leishmania spp. We evaluated the performance of the method using two distinct detection systems, such as SYBR Green and TaqMan probes, against a diverse panel of 225 clinical samples confirmed to have the disease. The real-time PCR targeting Hsp20 using SYBR Green demonstrated a sensitivity of 88% (95% CI: 83.53–92.47) and 100% specificity. Meanwhile, the TaqMan probe demonstrated a lower sensitivity of 47% (95% CI: 29.53–64.92). The high sensitivity and robust performance of the real-time PCR using SYBR Green establish its potential as a valuable diagnostic tool, particularly useful in endemic regions where rapid and accurate diagnosis is critical for timely treatment and effective disease control.

Instituciones: Universidad Nacional de San Cristóbal de Huamanga