A multiplex RT-PCR assay for the detection of chimeric transcripts encoded by the risk-stratifying translocations of pediatric acute lymphoblastic leukemia.
Modern therapy for pediatric acute lymphoblastic leukemia (ALL) is based on the principle of risk stratification. One of the most important laboratory features used to accurately risk stratify patients is the presence of specific chromosomal translocation within the leukemic blasts. In this paper, we describe a multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) assay for the accurate, sensitive, and rapid identification of chimeric transcripts encoded by the major risk-stratifying translocations of pediatric ALL. This assay will identify both the CML- and ALL-type BCR-ABL transcripts encoded by the t(9;22), all described variants of the E2A-PBX1 transcripts encoded by the t(1;19), the MLL-AF4 transcripts encoded by the t(4;11), and all variants of TEL-AML1 encoded by the t(12;21). In addition, we have developed a reverse dot-blot detection system as an alternative to traditional post-PCR Southern blot analysis. Application of this combined assay to the analysis of 70 leukemic samples and five cell lines resulted in a complete concordance between this multiplex assay and individual PCR reactions. The characteristics of the multiplex assay suggest that its application to routine clinical screening will significantly improve the ability of clinical laboratories to accurate risk stratify pediatric ALL patients.
Scurto
P
P
Department of Hematology/Oncology, St Jude Children's Research Hospital, and University of Tennessee College of Medicine, Memphis 38105, USA.
Hsu Rocha
M
M
Kane
J R
JR
Williams
W K
WK
Haney
D M
DM
Conn
W P
WP
Shurtleff
S A
SA
Downing
J R
JR
D002648:Child; D050676:Core Binding Factor Alpha 2 Subunit; D016044:Fusion Proteins, bcr-abl / Q000032:analysis; D018398:Homeodomain Proteins / Q000032:analysis; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000628:therapy; D051788:Myeloid-Lymphoid Leukemia Protein; D009363:Neoplasm Proteins / Q000032:analysis*; D015345:Oligonucleotide Probes / Q000235:genetics; D015514:Oncogene Proteins, Fusion / Q000032:analysis*; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics* / Q000628:therapy; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014178:Translocation, Genetic; D000097764:ETS Translocation Variant 6 Protein
D050676:Core Binding Factor Alpha 2 Subunit; D016044:Fusion Proteins, bcr-abl; D018398:Homeodomain Proteins; D051788:Myeloid-Lymphoid Leukemia Protein; D009363:Neoplasm Proteins; D015345:Oligonucleotide Probes; D015514:Oncogene Proteins, Fusion; C113855:MLL-AF4 fusion protein, human; C102421:TEL-AML1 fusion protein; D000097764:ETS Translocation Variant 6 Protein; C089847:E2A-Pbx1 fusion protein