Abstract / Summary
Leishmaniasis threatens nearly one billion people, with almost one million new cases annually across 98 countries; thus, rapid and accurate diagnosis is essential. Conventional microscopy and serology suffer from low sensitivity, invasive sampling, and an inability to separate active from past infection. Therefore, molecular tools are needed to improve detection and management. This systematic review and meta-analysis of thirty-three studies (2010–2026) evaluates recent advances in molecular diagnostics for human leishmaniasis, comparing accuracy, feasibility, and clinical relevance. We searched PubMed, Scopus, Web of Science, and Google Scholar. Pooled sensitivity and specificity were estimated using a bivariate random-effects model, and summary ROC curves were generated with the Hierarchical Summary Receiver Operating Characteristic (HSROC) model per Cochrane guidance. Heterogeneity was measured using the I² statistic. Estimates are reported with 95% confidence intervals. Quantitative real-time PCR (qPCR) targeting kinetoplast DNA (kDNA) minicircles achieved a pooled sensitivity of 95.8% (95% CI: 92.4–98.2%) and specificity of 98.8% (95% CI: 96.7–99.7%), serving as the reference standard despite high infrastructure needs. Loop-mediated isothermal amplification (LAMP) showed a pooled sensitivity of 92.5% (95% CI: 87.1–96.4%) and specificity of 98.9% (95% CI: 96.2–99.9%); however, heterogeneity was high (I² = 98.3%), mainly owing to primer mismatches for Leishmania aethiopica . Recombinase polymerase amplification (RPA) produced results in 15–20 min with strong agreement with qPCR (Cohen’s kappa = 0.89), supporting point-of-care use. Non-invasive samples, including urine, tape discs, and skin swabs, reached a pooled sensitivity of 93.9% (95% CI: 88.4–97.4%). Novel serology, such as BT7 plus EL1 peptides, achieved an AUC of 0.971, and recombinant Leishmania donovani Aurora kinase protein (LdAIRK) showed strong test-of-cure performance. Molecular methods have advanced; however, heterogeneity demands standardized protocols. Key gaps include the absence of next-generation sequencing (NGS) for direct diagnosis and a lack of cost-effectiveness analyses.