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Original Article


The 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

Vijay RamananKetki KelkarShatakshi RanadePriyanka GangodkarNikhita GogateKunal PatilTrupti Ragte-WathareMeenal AgarwalNikhil A. Phadke
MVR Foundation, Anjali Diagnostic Pathology Laboratory, Yashoda Hematology Clinic1GenePath Dx (Causeway Healthcare), Above Phadke Hospital, Pune, Maharashtra, India, 
Corresponding Author:

Nikhil D. Phadke

GenePath Dx (Causeway Healthcare), Above Phadke Hospital, Pune, Maharashtra, India
E-mail: nikhil.phadke@genepathdx.com

Corresponding Author:

Nikhil D. Phadke

GenePath Dx (Causeway Healthcare), Above Phadke Hospital, Pune, Maharashtra, India
E-mail: nikhil.phadke@genepathdx.com

DOI:10.18203/issn.2456-3994.IntJMolImmunoOncol20164386

ABSTRACT


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.
Keywords: NGS, multiplexing, CALR, JAK2, Hemato-oncology, Philadelphia-negative chronic myeloproliferative neoplasms