Research Papers:

A novel assay to detect calreticulin mutations in myeloproliferative neoplasms

Valentina Rosso, Jessica Petiti, Enrico Bracco, Roberto Pedrola, Francesca Carnuccio, Elisabetta Signorino, Sonia Carturan, Chiara Calabrese, Giada Bot-Sartor, Michela Ronconi, Anna Serra, Giuseppe Saglio, Francesco Frassoni and Daniela Cilloni _

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Oncotarget. 2017; 8:6399-6405. https://doi.org/10.18632/oncotarget.14113

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Valentina Rosso1, Jessica Petiti1, Enrico Bracco2, Roberto Pedrola1, Francesca Carnuccio1, Elisabetta Signorino1, Sonia Carturan1, Chiara Calabrese1, Giada Bot-Sartor1, Michela Ronconi1, Anna Serra1, Giuseppe Saglio1, Francesco Frassoni3, Daniela Cilloni1

1Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

2Department of Oncology, University of Turin, Turin, Italy

3Department of Pediatric Hemato-Oncology and Stem Cell, Cellular Therapy Laboratory, Institute G. Gaslini, Genova, Italy

Correspondence to:

Daniela Cilloni, email: [email protected]

Keywords: CALR, PNA, MPN, PCR clamping, diagnostic assay

Received: September 14, 2016     Accepted: December 15, 2016     Published: December 23, 2016


The myeloproliferative neoplasms are chronic myeloid cancers divided in Philadelphia positive (Ph+), chronic myeloid leukemia, or negative: polycythemia vera (PV) essential thrombocythemia (ET), and primary myelofibrosis (PMF). Most Ph negative cases have an activating JAK2 or MPL mutation. Recently, somatic mutations in the calreticulin gene (CALR) were detected in 56–88% of JAK2/MPL-negative patients affected by ET or PMF. The most frequent mutations in CARL gene are type-1 and 2. Currently, CALR mutations are evaluated by sanger sequencing. The evaluation of CARL mutations increases the diagnostic accuracy in patients without other molecular markers and could represent a new therapeutic target for molecular drugs.

We developed a novel detection assay in order to identify type-1 and 2 CALR mutations by PNA directed PCR clamping. Seventy-five patients affected by myeloproliferative neoplasms and seven controls were examined by direct DNA sequencing and by PNA directed PCR clamping. The assay resulted to be more sensitive, specific and cheaper than sanger sequencing and it could be applied even in laboratory not equipped for more sophisticated analysis. Interestingly, we report here a case carrying both type 1 and type2 mutations in CALR gene.

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