Enhanced blood parasite species identification using V4-V9 18S rDNA barcoding by universal primers on a nanopore platform.

ABSTRACT: Microscopic examination is commonly used for blood parasite detection in resource-limited settings due to its low cost and simplicity. However, it requires microscopy experts and has poor species-level identification. This study proposes a targeted next-generation sequencing (NGS) approach using a portable nanopore platform to enable accurate and sensitive parasite detection in such settings. To improve species identification on the error-prone nanopore sequencer, we designed a DNA barcoding strategy targeting the 18S rDNA V4-V9 region, which outperformed the commonly used V9 region. To enrich parasite DNA and reduce host contamination, two blocking primers were developed: a C3 spacer-modified oligo competing with the universal reverse primer and a peptide nucleic acid (PNA) oligo that inh

SUBMITTER: Sugi T 

PROVIDER: S-EPMC12639079 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature