Dataset Information

Flexible and cost-effective genomic surveillance of P. falciparum malaria with targeted nanopore sequencing.

ABSTRACT: Genomic surveillance of Plasmodium falciparum malaria can provide policy-relevant information about antimalarial drug resistance, diagnostic test failure, and the evolution of vaccine targets. Yet the large and low complexity genome of P. falciparum complicates the development of genomic methods, while resource constraints in malaria endemic regions can limit their deployment. Here, we demonstrate an approach for targeted nanopore sequencing of P. falciparum from dried blood spots (DBS) that enables cost-effective genomic surveillance of malaria in low-resource settings. We release software that facilitates flexible design of amplicon sequencing panels and use this software to design two target panels for P. falciparum. The panels generate 3-4 kbp reads for eight and sixteen targets respectively, covering key drug-resistance associated genes, diagnostic test antigens, polymorphic markers and the vaccine target csp. We validate our approach on mock and field samples, demonstrating robust sequencing coverage, accurate variant calls within coding sequences, the ability to explore P. falciparum within-sample diversity and to detect deletions underlying rapid diagnostic test failure.

SUBMITTER: de Cesare M

PROVIDER: S-EPMC10869361 | biostudies-literature | 2024 Feb

REPOSITORIES: biostudies-literature

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Flexible and cost-effective genomic surveillance of P. falciparum malaria with targeted nanopore sequencing.

de Cesare Mariateresa M Mwenda Mulenga M Jeffreys Anna E AE Chirwa Jacob J Drakeley Chris C Schneider Kammerle K Mambwe Brenda B Glanz Karolina K Ntalla Christina C Carrasquilla Manuela M Portugal Silvia S Verity Robert J RJ Bailey Jeffrey A JA Ghinai Isaac I Busby George B GB Hamainza Busiku B Hawela Moonga M Bridges Daniel J DJ Hendry Jason A JA

Nature communications 20240215 1

Genomic surveillance of Plasmodium falciparum malaria can provide policy-relevant information about antimalarial drug resistance, diagnostic test failure, and the evolution of vaccine targets. Yet the large and low complexity genome of P. falciparum complicates the development of genomic methods, while resource constraints in malaria endemic regions can limit their deployment. Here, we demonstrate an approach for targeted nanopore sequencing of P. falciparum from dried blood spots (DBS) that ena ...[more]

PMID: 38360754

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Project description:The genomic analysis of SARS-CoV-2 has served as a crucial tool for generating invaluable data that fulfils both epidemiological and clinical necessities. Long-read sequencing technology (e.g., ONT) has been widely used, providing a real-time and faster response when necessitated. A novel nanopore-based long-read sequencing platform named QNome nanopore has been successfully used for bacterial genome sequencing and assembly; however, its performance in the SARS-CoV-2 genomic surveillance is still lacking. Synthetic SARS-CoV-2 controls and 120 nasopharyngeal swab (NPS) samples that tested positive by real-time polymerase chain reaction were sequenced on both QNome and MGI platforms in parallel. The analytical performance of QNome was compared to the short-read sequencing on MGI. For the synthetic SARS-CoV-2 controls, despite the increased error rates observed in QNome nanopore sequencing reads, accurate consensus-level sequence determination was achieved with an average mapping quality score of approximately 60 (i.e., a mapping accuracy of 99.9999%). For the NPS samples, the average genomic coverage was 89.35% on the QNome nanopore platform compared with 90.39% for MGI. In addition, fewer consensus genomes from QNome were determined to be good by Nextclade compare with MGI (p < 0.05). A total of 9004 mutations were identified using QNome sequencing, with substitutions being the most prevalent, in contrast, 10 997 mutations were detected on MGI (p < 0.05). Furthermore, 23 large deletions (i.e., deletions≥ 10 bp) were identified by QNome while 19/23 were supported by evidence from short-read sequencing. Phylogenetic analysis revealed that the Pango lineage of consensus genomes for SARS-CoV-2 sequenced by QNome concorded 83.04% with MGI. QNome nanopore sequencing, though challenged by read quality and accuracy compared to MGI, is overcoming these issues through bioinformatics and computational advances. The advantage of structure variation (SV) detection capabilities and real-time data analysis renders it a promising alternative nanopore platform for the surveillance of the SARS-CoV-2.

Dataset Information

Flexible and cost-effective genomic surveillance of P. falciparum malaria with targeted nanopore sequencing.

Publications

Flexible and cost-effective genomic surveillance of P. falciparum malaria with targeted nanopore sequencing.

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