Multiplex ligation-dependent probe amplification for detection of genomic alterations in chronic lymphocytic leukaemia.
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.
Departament de Ciències Fisiològiques II, IDIBELL-Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain.
Llorenç
L
Coll-Mulet
Antonio F
AF
Santidrián
Ana M
AM
Cosialls
Daniel
D
Iglesias-Serret
Mercè
M
de Frias
Javier
J
Grau
Anna
A
Menoyo
Eva
E
González-Barca
Gabriel
G
Pons
Alicia
A
Domingo
Joan
J
Gil
D064007:Ataxia Telangiectasia Mutated Proteins; D018797:Cell Cycle Proteins; D002869:Chromosome Aberrations; D002872:Chromosome Deletion; D002881:Chromosomes, Human, Pair 12; D004268:DNA-Binding Proteins; D005784:Gene Amplification; D018628:Gene Dosage; D016158:Genes, p53; D023281:Genomics / Q000379:methods; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D017346:Protein Serine-Threonine Kinases; D013030:Spain; D025521:Tumor Suppressor Proteins
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D025521:Tumor Suppressor Proteins; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases