title: Multiplex ligation-dependent probe amplification and fluorescence in situ hybridization to detect chromosomal abnormalities in chronic lymphocytic leukemia: a comparative study.
abstract: Chronic lymphocytic leukemia (CLL) is a clinically heterogeneous disease characterized by recurrent chromosomal aberrations of prognostic significance. We aimed to evaluate the potential of the multiplex ligation-dependent probe amplification (MLPA) assay to detect genomic alterations in CLL. Highly purified (>90%) peripheral mononuclear CD19+ cell populations from 100 untreated CLL patients (pts) in early stage disease (Binet stage A) were included in this study. All samples were investigated by fluorescence in situ hybridization (FISH) for the presence of trisomy 12 and 17p13.1, 11q22.3, and 13q14.3 deletions. For MPLA analysis, DNA was amplified by means of two commercially available probes sets allowing the simultaneous screening of 56 genomic sequences. Overall, a high degree of concordance (95%) between MPLA and FISH results was found, if the abnormal clone was present in more than 30% of the leukemic cell population. The use of multiple MPLA probes allowed the fine-mapping of the 13q14 deletion and the identification of intragenic or small alterations undetected by FISH. Moreover, additional alterations in 2p24 (MYCN) (3 pts), 8q24 (MYC) (1 pt), 9p21 (CDKN2A2B) (1 pt), 1q21 (LMNA) (1 pt), and 6q25-26 (1 pt) regions not covered by a standard FISH assay were detected and all confirmed by FISH. Our data extend previously limited evidence that MLPA may represent a useful technique for the characterization of well-known lesions as well as the investigation of additional genomic changes in CLL.
authors.affiliation: Ematologia 1-CTMO, Fondazione Cà Granda IRCCS Policlinico and Dipartimento di Scienze Mediche, Università di Milano, Via F. Sforza 35, Milan, Italy.
authors.forename: Sonia
authors.initials: S
authors.lastname: Fabris
authors.forename: Oronzo
authors.initials: O
authors.lastname: Scarciolla
authors.forename: Fortunato
authors.initials: F
authors.lastname: Morabito
authors.forename: Rosa Anna
authors.initials: RA
authors.lastname: Cifarelli
authors.forename: Caterina
authors.initials: C
authors.lastname: Dininno
authors.forename: Giovanna
authors.initials: G
authors.lastname: Cutrona
authors.forename: Serena
authors.initials: S
authors.lastname: Matis
authors.forename: Anna Grazia
authors.initials: AG
authors.lastname: Recchia
authors.forename: Massimo
authors.initials: M
authors.lastname: Gentile
authors.forename: Gabriella
authors.initials: G
authors.lastname: Ciceri
authors.forename: Manlio
authors.initials: M
authors.lastname: Ferrarini
authors.forename: Angela
authors.initials: A
authors.lastname: Ciancio
authors.forename: Clara
authors.initials: C
authors.lastname: Mannarella
authors.forename: Antonino
authors.initials: A
authors.lastname: Neri
authors.forename: Alberto
authors.initials: A
authors.lastname: Fragasso
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018941:Antigens, CD19 / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics*; D002869:Chromosome Aberrations; D002872:Chromosome Deletion; D002880:Chromosomes, Human, Pair 11; D002881:Chromosomes, Human, Pair 12; D002882:Chromosomes, Human, Pair 13; D002886:Chromosomes, Human, Pair 17; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D011379:Prognosis; D017422:Sequence Analysis, DNA / Q000379:methods*; D014314:Trisomy
chemical_list: D018941:Antigens, CD19; D014408:Biomarkers, Tumor