Calreticulin Mutations in Myeloproliferative Neoplasms: Comparison of Three Diagnostic Methods.
Calreticulin (CALR) mutations have recently been reported in 70-84% of JAK2V617F-negative myeloproliferative neoplasms (MPN), and this detection has become necessary to improve the diagnosis of MPN. In a large single-centre cohort of 298 patients suffering from Essential Thrombocythemia (ET), the JAK2V617F, CALR and MPL mutations were noted in 179 (60%), 56 (18.5%) and 13 (4.5%) respectively. For the detection of the CALR mutations, three methods were compared in parallel: high-resolution melting-curve analysis (HRM), product-sizing analysis and Sanger sequencing. The sensitivity for the HRM, product-sizing analysis and Sanger sequencing was 96.4%, 98.2% and 89.3% respectively, whereas the specificity was 96.3%, 100% and 100%. In our cohort, the product-sizing analysis was the most sensitive method and was the easiest to interpret, while the HRM was sometimes difficult to interpret. In contrast, when large series of samples were tested, HRM provided results more quickly than did the other methods, which required more time. Finally, the sequencing method, which is the reference method, had the lowest sensitivity but can be used to describe the type of mutation precisely. Altogether, our results suggest that in routine laboratory practice, product-sizing analysis is globally similar to HRM for the detection of CALR mutations, and that both may be used as first-line screening tests. If the results are positive, Sanger sequencing can be used to confirm the mutation and to determine its type. Product-sizing analysis provides sensitive and specific results, moreover, with the quantitative measurement of CALR, which might be useful to monitor specific treatments.
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Ji-Hye
JH
Park
Inserm U866, Université de Bourgogne, Dijon, France.
Margaux
M
Sevin
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Selim
S
Ramla
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Aurélie
A
Truffot
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Tiffany
T
Verrier
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Dominique
D
Bouchot
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Martine
M
Courtois
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Mathilde
M
Bas
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Sonia
S
Benali
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
François
F
Bailly
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Bernardine
B
Favre
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Julien
J
Guy
Service d'anatomo-pathologie, CHU de Dijon, Dijon, France.
Laurent
L
Martin
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France.
Marc
M
Maynadié
UMR CNRS 5247, Université de Montpellier, Montpellier, France; Laboratoire de Cytologie Clinique, Pôle Biologie, CHU de Nîmes, Nîmes, France.
Serge
S
Carillo
Service d'Hématologie Biologique, Pôle Biologie, CHU de Dijon, Dijon, France; Inserm U866, Université de Bourgogne, Dijon, France; UMR CNRS 5247, Université de Montpellier, Montpellier, France; Laboratoire de Cytologie Clinique, Pôle Biologie, CHU de Nîmes, Nîmes, France; CRB Ferdinand Cabanne, Dijon, France.
François
F
Girodon
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D019046:Bone Marrow Neoplasms / Q000175:diagnosis* / Q000235:genetics; D037282:Calreticulin / Q000235:genetics*; D005260:Female; D016368:Frameshift Mutation / Q000235:genetics; D006801:Humans; D054643:INDEL Mutation / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009154:Mutation / Q000235:genetics; D012680:Sensitivity and Specificity; D017421:Sequence Analysis / Q000379:methods; D055815:Young Adult
D037282:Calreticulin