High resolution melting: a useful field-deployable method to measure dhfr and dhps drug resistance in both highly and lowly endemic Plasmodium populations

• Published in: Malaria Journal
• Main Authors: Yaye Dié Ndiaye, Cyrille K. Diédhiou, Amy K. Bei, Baba Dieye, Aminata Mbaye, Nasserdine Papa Mze, Rachel F. Daniels, Ibrahima M. Ndiaye, Awa B. Déme, Amy Gaye, Mouhamad Sy, Tolla Ndiaye, Aida S. Badiane, Mouhamadou Ndiaye, Zul Premji, Dyann F. Wirth, Souleymane Mboup, Donald Krogstad, Sarah K. Volkman, Ambroise D. Ahouidi, Daouda Ndiaye
• Format: Article in Journal/Newspaper
• Language: English
• Published: BMC 2017
• Subjects: Plasmodium falciparum; dhfr; dhps; HRM; PCR/RFLP; Senegal; Arctic medicine. Tropical medicine; RC955-962; Infectious and parasitic diseases; RC109-216; Arctic
• Online Access: https://doi.org/10.1186/s12936-017-1811-2; https://doaj.org/article/08790c5668104afeb7f2e4f6136d7d7d

Abstract Background Emergence and spread of drug resistance to every anti-malarial used to date, creates an urgent need for development of sensitive, specific and field-deployable molecular tools for detection and surveillance of validated drug resistance markers. Such tools would allow early detection of mutations in resistance loci. The aim of this study was to compare common population signatures and drug resistance marker frequencies between two populations with different levels of malaria endemicity and history of anti-malarial drug use: Tanzania and Sénégal. This was accomplished by implementing a high resolution melting assay to study molecular markers of drug resistance as compared to polymerase chain reaction–restriction fragment length polymorphism (PCR/RFLP) methodology. Methods Fifty blood samples were collected each from a lowly malaria endemic site (Sénégal), and a highly malaria endemic site (Tanzania) from patients presenting with uncomplicated Plasmodium falciparum malaria at clinic. Data representing the DHFR were derived using both PCR–RFLP and HRM assay; while genotyping data representing the DHPS were evaluated in Senegal and Tanzania using HRM. Msp genotyping analysis was used to characterize the multiplicity of infection in both countries. Results A high prevalence of samples harbouring mutant DHFR alleles was observed in both population using both genotyping techniques. HRM was better able to detect mixed alleles compared to PCR/RFLP for DHFR codon 51 in Tanzania; and only HRM was able to detect mixed infections from Senegal. A high prevalence of mutant alleles in DHFR (codons 51, 59, 108) and DHPS (codon 437) were found among samples from Sénégal while no mutations were observed at DHPS codons 540 and 581, from both countries. Overall, the frequency of samples harbouring either a single DHFR mutation (S108N) or double mutation in DHFR (C59R/S108N) was greater in Sénégal compared to Tanzania. Conclusion Here the results demonstrate that HRM is a rapid, sensitive, and field-deployable ...