Performance of High-Resolution Melting Analysis for Molecular Detection and Species Differentiation of Malaria: A Scoping Review

Malaria diagnosis remains challenging in low-density, asymptomatic, mixed-species, and non-falciparum infections. This scoping review mapped the diagnostic and operational evidence for high-resolution melting (HRM) analysis in malaria. Following PRISMA-ScR guidance, PubMed and Scopus were searched for English-language studies published between 2016 and 2026. Two reviewers independently screened records in Rayyan. Of 162 records identified, 15 studies met the eligibility criteria. Most assays targeted 18S rRNA, although mitochondrial, apicoplast, species-specific, and resistance-associated markers were also evaluated. Reported sensitivity ranged from 91.76% to 100%, and specificity generally exceeded 92%. HRM enabled differentiation of major human-infecting Plasmodium species and detected mixed, low-parasitemia, and submicroscopic infections that were missed by microscopy or rapid diagnostic tests. However, substantial heterogeneity in assay targets, platforms, comparators, and reporting limited direct comparison. Current applications remain laboratory-developed and require HRM-capable real-time PCR instruments, dedicated analytical software, and trained personnel. HRM is therefore a promising complementary method for reference laboratories and malaria surveillance, but standardized assays, multicentre validation, and implementation studies are required before routine or decentralized use.