RESUMO
Malaria remains the leading cause of morbidity and mortality in Chad. The World Health Organization (WHO) has recommended that endemic countries stratify malaria to guide interventions. Thus, the Republic of Chad has initiated a stratification process based on malaria incidence with the aim of defining transmission risk and proposing interventions. We collected routine malaria data from health facilities from 2017-2021, the national survey on malaria indicators, the entomological data of NMCP operational research, the demographic and health surveys, and remote sensing of environmental data. Stratification was based on the adjusted incidence of malaria to guide interventions. The adjusted incidence of malaria was, on average, 374 cases per 1000 people in the country. However, it varied according to health districts. Health districts were stratified into very low malaria incidence (n = 25), low malaria incidence (n = 20), moderate malaria incidence (n = 46) and high malaria incidence (n = 38). Micro-stratification in health districts with very low incidence was carried out to identify districts with incidence <10 cases per 1000 person with a view to a malaria pre-elimination programme. Appropriate malaria control interventions were proposed based on the strata identified. Stratification enables the country to target interventions to accelerate the reduction of the burden caused by malaria with a pre-elimination goal.
RESUMO
BACKGROUND: Given the risk of artemisinin resistance spreading from the Greater Mekong sub-region, prospective monitoring in sub-Saharan Africa should be expedited. Molecular biology techniques used for monitoring rely on the detection of k13 validated mutants by using PCR and Sanger sequencing approach, usually not available in malaria endemic areas. METHODS: A semi-automated workflow based on the easyMAG® platform and the Argene Solution® (bioMérieux, Marcy l'Etoile, France) as a field-based surveillance tool operable at national level was developed in four steps. Clinical and analytical performances of this tool detecting five of the most frequent and validated k13 mutants (Y493H, I543T, R539T, F446I and C580Y) from dried blood spots (DBS) were compared to the gold standard approach (PCR and Sanger sequencing). RESULTS: By using the ARMS (amplification-refractory mutation system) strategy, the best multiplexing options were found in 3 separate real-time PCR duplexes (IC as internal control/I543T, C580Y/Y493H and F446I/R539T) with limits of detection ranging from 50 (C580Y) to 6.25 parasites/µL (Y493H). In field conditions, using 642 clinical DBS (from symptomatic patients and asymptomatic individuals) collected from Cambodia, Myanmar and Africa (Chad), the overall sensitivity and specificity of the K13 bMx prototype assay developed by bioMérieux were ≥ 90%. Areas under the ROC curves were estimated to be > 0.90 for all k13 mutants in samples from symptomatic patients. CONCLUSION: The K13 ready-to-use bMx prototype assay, considered by the end-users as a user-friendly assay to perform (in shorter time than the K13 reference assay) and easy to interpret, was found to require less budget planning and had fewer logistical constraints. Its excellent performance qualifies the prototype as a reliable screening tool usable in malaria endemic countries recognized to be at risk of emergence or spread of validated k13 mutants. Additional multi-site studies are needed to evaluate the performances of the K13 bMx prototype assay in different epidemiological contexts such as Africa, India, or South America.
