TY - JOUR
T1 - Highly sensitive MYD88 l265p mutation detection by droplet digital polymerase chain reaction in waldenström macroglobulinemia
AU - Drandi, Daniela
AU - Genuardi, Elisa
AU - Dogliotti, Irene
AU - Ferrante, Martina
AU - Jiménez, Cristina
AU - Guerrini, Francesca
AU - Lo Schirico, Mariella
AU - Mantoan, Barbara
AU - Muccio, Vittorio
AU - Lia, Giuseppe
AU - Zaccaria, Gian Maria
AU - Omedè, Paola
AU - Passera, Roberto
AU - Orsucci, Lorella
AU - Benevolo, Giulia
AU - Cavallo, Federica
AU - Galimberti, Sara
AU - Sanz, Ramón García
AU - Boccadoro, Mario
AU - Ladetto, Marco
AU - Ferrero, Simone
N1 - Publisher Copyright:
© 2018 Ferrata Storti Foundation.
PY - 2018/6/3
Y1 - 2018/6/3
N2 - We here describe a novel method for MYD88 L265P mutation detection and minimal residual disease monitoring in Waldenström macroglobulinemia, by droplet digital polymerase chain reaction, in bone marrow and peripheral blood cells, as well as in circulating cell-free DNA. Our method shows a sensitivity of 5.00x10 -5 , which is far superior to the widely used allele-specific polymerase chain reaction (1.00x10 -3 ). Overall, 291 unsorted samples from 148 patients (133 with Waldenström macroglobulinemia, 11 with IgG lymphoplasmacytic lymphoma and 4 with IgM monoclonal gammopathy of undetermined sig-nificance) were analyzed: 194 were baseline samples and 97 were follow-up samples. One hundred and twenty-two of 128 (95.3%) bone marrow and 47/66 (71.2%) baseline peripheral blood samples scored positive for MYD88 L265P . To investigate whether MYD88 L265P detection by droplet digital polymerase chain reaction could be used for minimal residual disease monitoring, mutation levels were compared with IGH-based minimal residual disease analysis in 10 patients, and was found to be as informative as the classical, standardized, but not yet validated in Waldenström macroglobulinemia, IGH-based minimal residual disease assay (r 2 =0.64). Finally, MYD88 L265P detection by droplet digital polymerase chain reaction on plasma circulating tumor DNA from 60 patients showed a good correlation with bone marrow findings (bone marrow median mutational value 1.92x10 -2 , plasma circulating tumor DNA value: 1.4x10 -2 , peripheral blood value: 1.03x10 -3 ). This study indicates that droplet digital polymerase chain reaction assay of MYD88 L265P is a feasible and sensitive tool for mutation screening and minimal residual disease monitoring in Waldenström macroglobulinemia. Both unsorted bone marrow and peripheral blood samples can be reliably tested, as can circulating tumor DNA, which represents an attractive, less invasive alternative to bone marrow for MYD88 L265P detection.
AB - We here describe a novel method for MYD88 L265P mutation detection and minimal residual disease monitoring in Waldenström macroglobulinemia, by droplet digital polymerase chain reaction, in bone marrow and peripheral blood cells, as well as in circulating cell-free DNA. Our method shows a sensitivity of 5.00x10 -5 , which is far superior to the widely used allele-specific polymerase chain reaction (1.00x10 -3 ). Overall, 291 unsorted samples from 148 patients (133 with Waldenström macroglobulinemia, 11 with IgG lymphoplasmacytic lymphoma and 4 with IgM monoclonal gammopathy of undetermined sig-nificance) were analyzed: 194 were baseline samples and 97 were follow-up samples. One hundred and twenty-two of 128 (95.3%) bone marrow and 47/66 (71.2%) baseline peripheral blood samples scored positive for MYD88 L265P . To investigate whether MYD88 L265P detection by droplet digital polymerase chain reaction could be used for minimal residual disease monitoring, mutation levels were compared with IGH-based minimal residual disease analysis in 10 patients, and was found to be as informative as the classical, standardized, but not yet validated in Waldenström macroglobulinemia, IGH-based minimal residual disease assay (r 2 =0.64). Finally, MYD88 L265P detection by droplet digital polymerase chain reaction on plasma circulating tumor DNA from 60 patients showed a good correlation with bone marrow findings (bone marrow median mutational value 1.92x10 -2 , plasma circulating tumor DNA value: 1.4x10 -2 , peripheral blood value: 1.03x10 -3 ). This study indicates that droplet digital polymerase chain reaction assay of MYD88 L265P is a feasible and sensitive tool for mutation screening and minimal residual disease monitoring in Waldenström macroglobulinemia. Both unsorted bone marrow and peripheral blood samples can be reliably tested, as can circulating tumor DNA, which represents an attractive, less invasive alternative to bone marrow for MYD88 L265P detection.
UR - http://www.scopus.com/inward/record.url?scp=85048050213&partnerID=8YFLogxK
U2 - 10.3324/haematol.2017.186528
DO - 10.3324/haematol.2017.186528
M3 - Article
SN - 0390-6078
VL - 103
SP - 1029
EP - 1037
JO - Haematologica
JF - Haematologica
IS - 6
ER -