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Hence, there is need to gain a contemporary understanding of how indoor residual spraying (IRS) with non-pyrethroid insecticides when combined with long-lasting insecticidal nets (LLINs) impregnated with pyrethroid insecticides, contribute to the efforts of National Malaria Control Programmes to interrupt transmission and reduce the reservoir of <jats:italic>Plasmodium falciparum<\/jats:italic> infections across all ages. Using an interrupted time-series study design, four age-stratified malariometric surveys, each of ~2,000 participants, were undertaken pre- and post-IRS in Bongo District, Ghana. Following the application of three-rounds of IRS, <jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> transmission intensity declined, as measured by a &gt;90% reduction in the monthly entomological inoculation rate. This decline was accompanied by reductions in parasitological parameters, with participants of all ages being significantly less likely to harbor <jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> infections at the end of the wet season post-IRS (aOR = 0.22 [95% CI: 0.19\u20130.26], p-value &lt; 0.001). In addition, multiplicity of infection (MOI<jats:sub><jats:italic>var<\/jats:italic><\/jats:sub>) was measured using a parasite fingerprinting tool, designed to capture within-host genome diversity. At the end of the wet season post-IRS, the prevalence of multi-genome infections declined from 75.6% to 54.1%. This study demonstrates that in areas characterized by high seasonal malaria transmission, IRS in combination with LLINs can significantly reduce the reservoir of <jats:italic>P<\/jats:italic>. <jats:italic>falciparum<\/jats:italic> infection. Nonetheless despite this success, 41.6% of the population, especially older children and adolescents, still harboured multi-genome infections. Given the persistence of this diverse reservoir across all ages, these data highlight the importance of sustaining vector control in combination with targeted chemotherapy to move high-transmission settings towards pre-elimination. This study also points to the benefits of molecular surveillance to ensure that incremental achievements are not lost and that the goals advocated for in the WHO\u2019s High Burden to High Impact strategy are realized.<\/jats:p>","DOI":"10.1371\/journal.pgph.0000285","type":"journal-article","created":{"date-parts":[[2022,5,18]],"date-time":"2022-05-18T17:25:31Z","timestamp":1652894731000},"page":"e0000285","update-policy":"https:\/\/doi.org\/10.1371\/journal.pgph.corrections_policy","source":"Crossref","is-referenced-by-count":38,"title":["Indoor residual spraying with a non-pyrethroid insecticide reduces the reservoir of Plasmodium falciparum in a high-transmission area in northern Ghana"],"prefix":"10.1371","volume":"2","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3305-0533","authenticated-orcid":true,"given":"Kathryn E.","family":"Tiedje","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abraham 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