title: Evaluation of multiplex ligation-dependent probe amplification as a method for the detection of copy number abnormalities in B-cell precursor acute lymphoblastic leukemia.
abstract: Recent genomic studies have shown that copy number abnormalities (CNA) of genes involved in lymphoid differentiation and cell cycle control are common in B-cell precursor acute lymphoblastic leukemia (BCP-ALL). We have evaluated Multiplex Ligation-dependent Probe Amplification (MLPA) on 43 BCP-ALL patients for the detection of the most common deletions among these genes and compared the results to those obtained by fluorescence in situ hybridization (FISH) and genomic quantitative PCR (qPCR). There was good correlation between methods for CDKN2A/B, IKZF1, and PAX5 deletions in the majority of cases and MLPA confirmed the presence of deletions within the PAR1 region in two of three cases identified by FISH. Small intragenic aberrations detected by MLPA, which were below the resolution of FISH for CDKN2A/B (n = 7), IKZF1 (n = 3), and PAX5 (n = 3) were confirmed by qPCR. MLPA and qPCR were unable to detect populations present at a low level (<20%) by FISH. In addition, although MLPA identified the presence of a deletion, it was unable to discern the presence of mixed cell populations which had been identified by FISH: CDKN2A/B (n = 3), IKZF1 (n = 1), PAX5 (n = 2), and PAR1 deletion (n = 1). Nevertheless, this study has demonstrated that MLPA is a robust technique for the reliable detection of CNA involving multiple targets in a single test and thus is ideal for rapid high throughput testing of large cohorts with a view to establishing incidence and prognostic significance.
authors.affiliation: Leukaemia Research Cytogenetics Group, Northern Institute for Cancer Research, Newcastle University, Royal Victoria Infirmary, Newcastle upon Tyne, UK.
authors.forename: C J
authors.initials: CJ
authors.lastname: Schwab
authors.forename: L R
authors.initials: LR
authors.lastname: Jones
authors.forename: H
authors.initials: H
authors.lastname: Morrison
authors.forename: S L
authors.initials: SL
authors.lastname: Ryan
authors.forename: H
authors.initials: H
authors.lastname: Yigittop
authors.forename: J P
authors.initials: JP
authors.lastname: Schouten
authors.forename: C J
authors.initials: CJ
authors.lastname: Harrison
mesh_terms: D002453:Cell Cycle; D055028:Comparative Genomic Hybridization / Q000379:methods; D020732:Cytogenetic Analysis / Q000379:methods*; D056915:DNA Copy Number Variations; D015342:DNA Probes; D018628:Gene Dosage; D018816:Genes, cdc; D019942:Genes, p16; D006801:Humans; D051740:Ikaros Transcription Factor / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D007621:Karyotyping / Q000379:methods; D008214:Lymphocytes; D015336:Molecular Probe Techniques; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D051757:PAX5 Transcription Factor / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D015452:Precursor B-Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics*; D012680:Sensitivity and Specificity; D014157:Transcription Factors / Q000235:genetics
chemical_list: D015342:DNA Probes; D051757:PAX5 Transcription Factor; C497281:PAX5 protein, human; D014157:Transcription Factors; D051740:Ikaros Transcription Factor