Open AccessThe clinical utility of a custom-developed targeted next-generation sequencing assay for detection of mutations associated with Philadelphia-negative chronic myeloproliferative neoplasms: Two case examples with CALR exon 9 mutations
Author(s) -
Vijay Ramanan,
Ketki Kelkar,
Shatakshi Ranade,
Priyanka Gangodkar,
Nikhita Gogate,
Kunal Patil,
Trupti Ragte-Wathare,
Meenal Agarwal,
Nikhil Phadke
Publication year - 2016
Publication title -
international journal of molecular and immuno oncology
Language(s) - English
Resource type - Journals
eISSN - 2836-3779
pISSN - 2456-3994
DOI - 10.18203/issn.2456-3994.intjmolimmunooncol20164386
Subject(s) - exon , mutation , dna sequencing , genetics , biology , cancer research , myeloproliferative disorders , gene , immunology
We have developed a highly targeted custom next generation sequencing-based test targeting JAK2 exons 12 and 14, CALR exon 9, and MPL exon 10, which are implicated in Philadelphia-negative chronic myeloproliferative neoplasms. The assay is capable of ultra-high sequencing depth and producing mutational detection sensitivities of 0.5% and below. We have validated the performance of the test through orthogonal testing and demonstrated a high degree of multiplexing with up to 50 samples in a single run. We show the clinical utility of this test through the description of a couple of cases with myeloproliferative disorders with Type-II and Type-II-like mutations in exon 9 of the CALR gene.