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Blood, 1 February 2008, Vol. 111, No. 3, pp. 1124-1127.
Prepublished online as a Blood First Edition Paper on November 13, 2007; DOI 10.1182/blood-2007-06-093302.


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Submitted June 6, 2007
Accepted October 28, 2007

Development of an allele-specific minimal residual disease assay for patients with juvenile myelomonocytic leukemia

Sophie Archambeault, Nikki J Flores, Ayami Yoshimi, Christian P. Kratz, Miriam Reising, Alexandra Fischer, Peter Noellke, Franco Locatelli, Petr Sedlacek, Christian Flotho, Marco Zecca, Peter D. Emanuel, Robert P. Castleberry, Charlotte M Niemeyer, Peter Bader, and Mignon L. Loh*

Department of Pediatrics, University of California, San Francisco, CA, United States
European Working Group of Myelodysplastic Syndromes in Childhood
Children's Oncology Group, Arcadia, CA, United States
Comprehensive Cancer Center, University of California, San Francisco, CA, United States

* Corresponding author; email: lohm{at}peds.ucsf.edu.

Juvenile myelomonocytic leukemia is an aggressive and frequently lethal myeloproliferative disorder of childhood. Somatic mutations in NRAS, KRAS, or PTPN11 occur in 60% of cases. Monitoring disease status is difficult due to the lack of characteristic leukemic blasts at diagnosis. We designed a fluorescently based, allele-specific polymerase chain reaction assay called TaqMAMA to detect the most common RAS, or PTPN11 mutations. We analyzed peripheral blood and/or bone marrow of 25 patients for levels of mutant alleles over time. Analysis of pre-hematopoietic stem cell transplant (HSCT) samples revealed a broad distribution of the quantity of the mutant alleles. After HSCT, the level of the mutant allele rose rapidly in patients who relapsed and correlated well with falling donor chimerism. Simultaneously analyzed peripheral blood and bone marrow samples demonstrate that blood can be monitored for residual disease. Importantly, these assays provide a sensitive strategy to evaluate molecular responses to new therapeutic strategies.


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