<p>In sub-Saharan Africa, continental-scale genomic surveillance of <i>Plasmodium falciparum</i> malaria is needed to track the spread of drug and diagnostic resistance, as well as monitor parasite evolutionary responses to vaccine rollout. Yet continental-scale implementation is hindered by a lack of genomic approaches suitable for local laboratories, and the vastness of the continent. Here, we initiate a decentralised scale-up of <i>P. falciparum</i> genomic surveillance by locally sequencing and analysing 1065 samples across six African countries in one year. We achieve this with a novel nanopore sequencing protocol that is rapid (~5 hr) and cost-effective (&lt;$25 USD/sample), providing surveillance of antimalarial drug resistance genes, <i>hrp2/3</i> deletions, the vaccine target <i>csp</i>, and the polymorphic gene <i>ama1</i>. We couple this to a laptop-based bioinformatics dashboard that runs offline and displays mapping and variant calling results in real-time. We demonstrate robust sequencing coverage across parasitemia levels and laboratories, accurate identification of antimalarial resistance markers and <i>hrp2/3</i> deletions; and, with a novel variant caller, sensitive detection of mutations carried by minor clones. Our approach will accelerate genomic surveillance of <i>P. falciparum</i> malaria across sub-Saharan Africa at a time of urgent need.</p>

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Continental-scale genomic surveillance of Plasmodium falciparum malaria across sub-Saharan Africa with rapid nanopore sequencing

  • Mulenga Mwenda,
  • Karolina Mosler,
  • Bernd Bohmeier,
  • Miriam Chomba,
  • Welmoed van Loon,
  • Brenda Mambwe,
  • Amy Gaye,
  • Adedolapo Olorunfemi,
  • Salma Suliman,
  • Nassandba Julien Yanogo,
  • Djiby Sow,
  • Bassirou Ngom,
  • Oumou Aïcha Zeïna Zoure,
  • Yssimini Nadège Guillène Tibiri,
  • Fiyinfoluwa Ojeniyi,
  • Arsène Zongo,
  • Etilé A. Anoh,
  • Vincent Achi,
  • Carol Chiyesu,
  • Sheila Otieno,
  • Bixa Ogola,
  • Moussa Niangaly,
  • Manuela Carrasquilla,
  • Dagaga Kenea Goboto,
  • Torsten Feldt,
  • Tafese Beyene Tufa,
  • Rafael Oliveira,
  • Emma Schallenberg,
  • Yuhana Sogoba,
  • Christina Ntalla,
  • Oumarou Ouedraogo,
  • Kephas Otieno,
  • Oluyinka Opaleye,
  • Adekunle Olowe,
  • Marley Gibbons,
  • Chris Drakeley,
  • Grit Schubert,
  • Frank P. Mockenhaupt,
  • Silvia Portugal,
  • Awa B. Deme,
  • Issiaka Soulama,
  • Daouda Ndiaye,
  • Olusola Ojurongbe,
  • Simon Kariuki,
  • Ya Ping Shi,
  • Jonathan S. Schultz,
  • Moonga Hawela,
  • Daniel J. Bridges,
  • Jason A. Hendry

摘要

In sub-Saharan Africa, continental-scale genomic surveillance of Plasmodium falciparum malaria is needed to track the spread of drug and diagnostic resistance, as well as monitor parasite evolutionary responses to vaccine rollout. Yet continental-scale implementation is hindered by a lack of genomic approaches suitable for local laboratories, and the vastness of the continent. Here, we initiate a decentralised scale-up of P. falciparum genomic surveillance by locally sequencing and analysing 1065 samples across six African countries in one year. We achieve this with a novel nanopore sequencing protocol that is rapid (~5 hr) and cost-effective (<$25 USD/sample), providing surveillance of antimalarial drug resistance genes, hrp2/3 deletions, the vaccine target csp, and the polymorphic gene ama1. We couple this to a laptop-based bioinformatics dashboard that runs offline and displays mapping and variant calling results in real-time. We demonstrate robust sequencing coverage across parasitemia levels and laboratories, accurate identification of antimalarial resistance markers and hrp2/3 deletions; and, with a novel variant caller, sensitive detection of mutations carried by minor clones. Our approach will accelerate genomic surveillance of P. falciparum malaria across sub-Saharan Africa at a time of urgent need.