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Updated: Sep 19, 2026
Sequencing of mRNA from Whole Blood using Nanopore Sequencing
Published on: June 3, 2019
Nanopore-based, long-range Parvovirus B19 amplicon sequencing for near-whole genome characterization
C Akile1, B M Hogema2, M W A Molenaar-de Backer3
1Department of Medical Microbiology and Infection Control, Leiden University Center of Infectious Diseases, Leiden University Medical Center, Leiden, the Netherlands.
Background:
Whole-Genome Sequencing (WGS) enables monitoring of genomic variation and evaluation of diagnostic PCR assays. However, WGS data for Parvovirus B19 (B19V) remains limited despite its relevance for clinical care and transfusion safety. To increase the availability of high-quality B19V genomic data, a near-WGS protocol was developed and validated.
Method:
The protocol combines long-range PCR to generate a 4.6-kb amplicon, covering ∼82% of the B19V genome, with Oxford Nanopore sequencing. Validation was performed using six reference samples and nineteen B19V-positive donor plasma samples.
Results:
After quality control, samples achieved a median sequencing depth of 152x. Sequences generated from the six reference samples showed 100% concordance with previously published data. Genomic analysis of donor samples explained atypical amplification profiles observed during routine PCR screening.
Conclusion:
The newly developed protocol provides a scalable method for B19V genome characterization, enabling assessment of oligonucleotide-binding regions for PCR assay monitoring and facilitating the generation of genomic data for future epidemiological investigations.
