Evaluation of methods to detect CALR mutations in myeloproliferative neoplasms
| Autor: | Alison Callaway, Carolyn L. Dent, Jyoti Nangalia, Amy V. Jones, Helen E. White, Chris Mattocks, Daniel Ward, Nicholas C.P. Cross, Hans G. Drexler, Matthew Lyon, Andrew Chase, William T. M. Leung |
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| Rok vydání: | 2015 |
| Předmět: |
Male
Cancer Research DNA Mutational Analysis Biology medicine.disease_cause DNA sequencing High Resolution Melt symbols.namesake Exon chemistry.chemical_compound Screening method medicine Humans Indel Alleles Sanger sequencing Genetics Mutation Myeloproliferative Disorders DNA Neoplasm Hematology Neoplasm Proteins Oncology chemistry Hematologic Neoplasms symbols Female Lod Score Calreticulin DNA |
| Zdroj: | Leukemia Research. 39:82-87 |
| ISSN: | 0145-2126 |
| DOI: | 10.1016/j.leukres.2014.11.019 |
| Popis: | The recent discovery of somatically acquired CALR mutations in a substantial proportion of patients with myeloproliferative neoplasms has provided a new marker of clonal disease, advancing both diagnosis and prognosis in these previously difficult to characterise disorders. The mutations, which can be challenging to detect on a routine basis, are heterogeneous insertions/deletions (indels) in exon 9 with mutant allele burden that vary substantially between patients. We evaluated four genetic screening methods for their ability to detect a series of different CALR mutations; Sanger sequencing, fragment analysis PCR, high resolution melt (HRM) and targeted next generation sequencing (NGS). The limit of detection (LoD) of each assay was tested using serial dilution series made with DNA from CALR positive sample DNA and a cell line, MARIMO, found to carry a heterozygous 61 nucleotide CALR deletion. All methods were capable of detecting each mutation; HRM and fragment analysis PCR were better at detecting low mutation levels compared to Sanger sequencing but targeted NGS had the lowest LoD at a 1% mutation burden. |
| Databáze: | OpenAIRE |
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