Multiplex ligation‐dependent probe amplification for detection of genomic alterations in chronic lymphocytic leukaemia

Abstract

Summary Chronic lymphocytic leukaemia (CLL) is the commonest form of leukaemia in adults in Western countries. We performed multiplex ligation‐dependent probe amplification (MLPA) analysis in 50 CLL patients to identify multiple genomic CLL‐specific targets, including genes located at 13q14, 17p13 (TP53), 11q23 (ATM) and chromosome 12, and compared the results with those obtained with fluorescence in situ hybridization (FISH). There was a good correlation between MLPA and FISH results, as most alterations (89%) were detected by both techniques. Only three cases with a low percentage (<25%) of cells carrying the alterations were not detected by MLPA. On the other hand, as MLPA uses multiple probes it identified intragenic or small alterations undetected by FISH in three cases. MLPA also detected alterations in 8q24 (MYC) and 6q25–26. In summary, unlike interphase FISH, MLPA enabled the simultaneous analysis of many samples with automated data processing at a low cost. Therefore, the combination of robust multiplexing and high throughput makes MLPA a useful technique for the analysis of genomic alterations in CLL.

AICAR induces apoptosis independently of AMPK and p53 through up-regulation of the BH3-only proteins BIM and NOXA in chronic lymphocytic leukemia cells

“…15 Patient 26 has a 17p13 deletion in one allele in 43% of peripheral blood lymphocytes, and patient 27 has a 17p13 deletion in one allele in 94% of peripheral blood lymphocytes. 17 Samples from patients 7 and 28 have 11q22 deletion and do not express ATM protein. 17,18 Patient 7 has the 11q23 deletion in one allele in 86% of peripheral blood lymphocytes, and patient 29 has the 11q23 deletion determined by genomic multiplex ligation-dependent probe amplification (MLPA; relative copy number value ϭ 0.57).…”

Akt inhibitors induce apoptosis in chronic lymphocytic leukemia cells

“…Patients' samples with deleted/mutated TP53 or altered expression have been described previously. [28][29][30] Patient 8 had a 17p deletion in one allele in 43% of peripheral blood lymphocytes, patient 41 had a 17p deletion in one allele in 94% of peripheral blood lymphocytes, and patient 40 had a frame-shift mutation in one allele (nucleotide deletion in codon 272) and a 17p deletion in the other allele in 86% of peripheral blood lymphocytes. First, we incubated these CLL cells with 5 µM Akti-1/2 or 0.5 µM A-443654 for 24 h. Interestingly, we observed a decrease in viability in two of the three samples with Akti-1/2 treatment and in all three samples with A-443654 treatment ( Figure 6A).…”

Prognostification of ALL by Cytogenetics

“…Although MLPA is not suitable for genome-wide research screening, it provides a low cost, simple alternative to array-based techniques for many routine applications. It is now used routinely to identify copy number alterations in haematological malignancies for which specific copy number changes are clinically significant [6].…”