Association of clinical status of follicular lymphoma patients after autologous stem cell transplant and quantitative assessment of lymphoma in blood and bone marrow as measured by SYBR Green I polymerase chain reaction.
Molecular remission in the autograft and bone marrow after transplant are predictive of durable clinical remission in relapsed follicular lymphoma. Thus, a simple reliable method to quantify minimal residual disease (MRD) would improve prognostication in these patients. Fluorescent hybridization probes have been used in real-time quantitative polymerase chain reaction (RQ-PCR) to monitor MRD with a reproducible sensitivity of 0.01%; however, these techniques are expensive and require additional experiments to examine clonality. We describe a SYBR Green I detection method that is more universal, checks clonal identity, yields the same sensitivity for monitoring MRD, and is more economically attractive. Using this method to follow 14 follicular lymphoma patients treated with autologous stem cell transplantation, molecular markers were successfully defined for 12 patients. Median contamination of stem-cell grafts was 0.1% (range, 0 to 13%). Six patients with measurable graft contamination became PCR-negative in blood and bone marrow within 12 months after autologous stem cell transplantation. Three patients free of disease progression (median follow-up of 75 months) are in molecular remission. Increasing fractions of RQ-PCR-positive blood and bone marrow cells reliably predicted morphological and clinical relapse. In one case, both clinical relapse and spontaneous regression were reflected by changes in MRD levels. Thus, our RQ-PCR method reproducibly distinguishes different levels of MRD.
Pennell
Nancy
N
Advanced Therapeutics Program, Toronto-Sunnybrook Regional Cancer Centre, Sunnybrook and Women's College Health Sciences Centre, 2075 Bayview Ave., Toronto, ON Canada M4N 3M5.
Woods
Anthony
A
Reis
Marciano
M
Buckstein
Rena
R
Spaner
David
D
Imrie
Kevin
K
Hewitt
Karen
K
Boudreau
Angela
A
Seth
Arun
A
Berinstein
Neil L
NL
D000293:Adolescent; D000328:Adult; D052160:Benzothiazoles; D014408:Biomarkers, Tumor / Q000032:analysis; D001853:Bone Marrow / Q000473:pathology; D003959:Diamines; D005456:Fluorescent Dyes; D006801:Humans; D008224:Lymphoma, Follicular / Q000175:diagnosis* / Q000473:pathology / Q000628:therapy; D008875:Middle Aged; D018365:Neoplasm, Residual / Q000175:diagnosis; D009930:Organic Chemicals; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D012680:Sensitivity and Specificity; D033581:Stem Cell Transplantation; D014182:Transplantation, Autologous
D052160:Benzothiazoles; D014408:Biomarkers, Tumor; D003959:Diamines; D005456:Fluorescent Dyes; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I