A diagnostic biochip for the comprehensive analysis of MLL translocations in acute leukemia.
Reciprocal rearrangements of the MLL gene are among the most common chromosomal abnormalities in both Acute Lymphoblastic and Myeloid Leukemia. The MLL gene, located on the 11q23 chromosomal band, is involved in more than 40 recurrent translocations. In the present study, we describe the development and validation of a biochip-based assay designed to provide a comprehensive molecular analysis of MLL rearrangements when used in a standard clinical pathology laboratory. A retrospective blind study was run with cell lines (n=5), and MLL positive and negative patient samples (n=31), to evaluate assay performance. The limits of detection determined on cell line data were 10(-1), and the precision studies yielded 100% repeatability and 98% reproducibility. The study shows that the device can detect frequent (AF4, AF6, AF10, ELL or ENL) as well as rare partner genes (AF17, MSF). The identified fusion transcripts can then be used as molecular phenotypic markers of disease for the precise evaluation of minimal residual disease by RQ-PCR. This biochip-based molecular diagnostic tool allows, in a single experiment, rapid and accurate identification of MLL gene rearrangements among 32 different fusion gene (FG) partners, precise breakpoint positioning and comprehensive screening of all currently characterized MLL FGs.
IPSOGEN SAS, Case 923, 163, Av. de Luminy, Marseille Cedex 9, France. maroc@ipsogen.com
N
N
Maroc
A
A
Morel
E
E
Beillard
A L
AL
De La Chapelle
X
X
Fund
M-J
MJ
Mozziconacci
M
M
Dupont
J-M
JM
Cayuela
J
J
Gabert
A
A
Koki
V
V
Fert
F
F
Hermitte
D000208:Acute Disease; D014408:Biomarkers, Tumor / Q000235:genetics*; D045744:Cell Line, Tumor; D002880:Chromosomes, Human, Pair 11 / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015321:Gene Rearrangement; D011495:Histone-Lysine N-Methyltransferase; D006801:Humans; D007938:Leukemia / Q000175:diagnosis / Q000235:genetics*; D051788:Myeloid-Lymphoid Leukemia Protein; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods*; D015514:Oncogene Proteins, Fusion / Q000032:analysis / Q000235:genetics*; D011519:Proto-Oncogenes; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D014157:Transcription Factors; D014178:Translocation, Genetic; D016335:Zinc Fingers
D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; C497422:KMT2A protein, human; D015514:Oncogene Proteins, Fusion; D012334:RNA, Neoplasm; D014157:Transcription Factors; D051788:Myeloid-Lymphoid Leukemia Protein; D011495:Histone-Lysine N-Methyltransferase