Validation of polymerase chain reaction-indexed next-generation sequencing assay on the Illumina iSeq 100 platform for MPL exon 10 mutation detection: a robust alternative to Sanger sequencing

Myeloproliferative neoplasms (MPNs) are clonal disorders characterised by excessive proliferation of myeloid lineages. Mutations in MPL exon 10, including p.Trp515Leu and p.Trp515Lys, are recurrent and clinically relevant. 1 2 Sanger sequencing is traditionally used for DNA sequencing3 but has limited sensitivity, typically detecting variants at approximately 15%–20% variant allele frequency (VAF) in routine practice.4