|
Title
|
|
|
|
Selective whole-genome sequencing of Plasmodium parasites directly from blood samples by nanopore adaptive sampling
| |
|
Author
|
|
|
|
| |
|
Abstract
|
|
|
|
| Whole-genome sequencing of Plasmodium is becoming an increasingly important tool for genomic surveillance of malaria. Due to the predominance of human DNA in a patient blood sample, time-consuming laboratory procedures are required to deplete human DNA or enrich Plasmodium DNA. Here, we investigated the potential of nanopore adaptive sampling to enrich Plasmodium falciparum reads while sequencing unenriched patient blood samples. To compare adaptive sampling versus regular sequencing on a MinION device, a dilution series consisting of 0%–84% P . falciparum DNA in human DNA was sequenced. Half of the flow cell channels were run in adaptive sampling mode, enriching for the P. falciparum reference genome, resulting in a three- to five-fold enrichment of P. falciparum bases in samples containing 0.1%–8.4% P. falciparum DNA. This finding was confirmed by sequencing three P. falciparum patient blood samples with common levels of parasitemia, that is, 0.1%, 0.2%, and 0.6% in adaptive mode. Their estimated enrichment was 5.8, 3.9, and 2.7, respectively, which was sufficient to cover at least 97% of the P. falciparum reference genome at a median depth of 5 (lowest parasitemia) or 355 (highest parasitemia). In all, 38 drug resistance loci were compared to Sanger sequencing results, showing high concordance, which suggests that the obtained sequencing data are of sufficient quality to address common clinical research questions for patients with parasitemias of 0.1% and higher. Overall, our results indicate that adaptive nanopore sequencing has the potential to replace more time-consuming Plasmodium enrichment protocols in the future. |
| |
|
Language
|
|
|
|
English
| |
|
Source (journal)
|
|
|
|
mBio / American Society for Microbiology. - Washington, D.C., 2010, currens
| |
|
Publication
|
|
|
|
Washington, D.C.
:
American Society for Microbiology
,
2024
| |
|
ISSN
|
|
|
|
2150-7511
| |
|
DOI
|
|
|
|
10.1128/MBIO.01967-23
| |
|
Volume/pages
|
|
|
|
15
:1
(2024)
, p. 1-15
| |
|
Article Reference
|
|
|
|
e01967-23
| |
|
ISI
|
|
|
|
001114831100001
| |
|
Pubmed ID
|
|
|
|
38054750
| |
|
Full text (Publisher's DOI)
|
|
|
|
| |
|
Full text (open access)
|
|
|
|
| |
|