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Comprehensive chronic lymphocytic leukemia diagnostics by combined multiplex ligation dependent probe amplification (MLPA) and interphase fluorescence in situ hybridization (iFISH) (CROSBI ID 227180)

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Alhourani, Eyad ; Rincic, Martina ; Othman, Moneeb AK ; Pohle, Beate ; Schlie, Cordula ; Glaser, Anita ; Liehr, Thomas Comprehensive chronic lymphocytic leukemia diagnostics by combined multiplex ligation dependent probe amplification (MLPA) and interphase fluorescence in situ hybridization (iFISH) // Molecular Cytogenetics, 7 (2014),

Podaci o odgovornosti

Alhourani, Eyad ; Rincic, Martina ; Othman, Moneeb AK ; Pohle, Beate ; Schlie, Cordula ; Glaser, Anita ; Liehr, Thomas

engleski

Comprehensive chronic lymphocytic leukemia diagnostics by combined multiplex ligation dependent probe amplification (MLPA) and interphase fluorescence in situ hybridization (iFISH)

Background: Banding-karyotyping and metaphase-directed-fluorescence-in-situhybridization (FISH) may be hampered by low mitotic index in leukemia. Interphase FISH (iFISH) is a way out here, however, testing many probes at the same time is protracted and expensive. Here multiplex-ligation-dependent-probe-amplification (MLPA) was used retrospectively in chronic lymphocytic leukemia (CLL) samples initially studied by banding cytogenetics and iFISH. Detection rates of iFISH and MLPA were compared and thus a cost-efficient scheme for routine diagnostics is proposed. Results: Banding cytogenetics was done successfully in 67/85 samples. DNA was extracted from all 85 CLL samples. A commercially available MLPA probe set directed against 37 loci prone to be affected in hematological malignancies was applied. Besides, routine iFISH was done by commercially available probes for following regions: 11q22.3, 12p11.2-q11.1, 13q14.3, 13q34, 14q32.33 and 17p13.1. MLPA results were substantiated by iFISH using corresponding locus-specific probes. Aberrations were detected in 67 of 85 samples (similar to 79%) applying banding cytogenetics, iFISH and MLPA. A maximum of 8 aberrations was detected per sample ; however, one aberration per sample was found most frequently. Overall 163 aberrations were identified. 15 of those (similar to 9%) were exclusively detected by banding cytogenetics, 95 were found by MLPA (similar to 58%) and 100 (similar to 61%) by routine iFISH. MLPA was not able to distinguish reliably between mono-and biallelic del(13)(q14.3q14.3), which could be easily identified as well as quantified by routine iFISH. Also iFISH was superior to MLPA in samples with low tumor cell load. On the other hand MLPA detected additional aberrations in 22 samples, two of them being without any findings after routine iFISH. Conclusions: Both MLPA and routine iFISH have comparable detection rates for aberrations being typically present in CLL. As MLPA can detect also rare chromosomal aberrations it should be used as an initial test if routine cytogenetics is not possible or non-informative. Still iFISH should be used additionally to distinguish mono-from biallelic deletions and also to determine rate of mosaicism for 13q14.2 to 13q14.3. In case MLPA is negative the corresponding CLL samples should be tested at least by iFISH using the standard probe set to.

Chronic lymphocytic leukemia (CLL); Chromosomal aberrations; Multiplex ligation-dependent probe amplification (MLPA); Fluorescence in situ hybridization (FISH)

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Podaci o izdanju

7

2014.

objavljeno

1755-8166

Povezanost rada

Temeljne medicinske znanosti

Indeksiranost