PMID- 12679597
OWN - NLM
STAT- MEDLINE
DCOM- 20031105
LR  - 20171101
IS  - 1015-2008 (Print)
IS  - 1015-2008 (Linking)
VI  - 70
IP  - 4
DP  - 2002-2003
TI  - Significant hybridization differences in comparative genomic hybridization due to 
      nucleotides used for DNA labelling and to DNA chosen for cohybridization.
PG  - 204-8
AB  - OBJECTIVE: Comparative genomic hybridization (CGH) has been established as an 
      informative technique in genetic analysis. However, differences in the ratio of 
      hybridization intensities were reported for particular chromosomes, which may 
      affect CGH results. The aim of this study was to define these differences in more 
      detail. For this purpose, CGH results of 70 samples of bone marrow cells (BMC) 
      with normal karyotype in conventional cytogenetics (CC) were evaluated using 
      seven different reference DNAs and two different DNA labeling systems. METHODS 
      AND RESULTS: CGH using fluorochrome-conjugated nucleotides for DNA labeling 
      indicated signal deviations in 21/70 BMC samples. Deviations affected chromosomes 
      1 (n = 21), 2 (n = 11), 4 (n = 11), 5 (n = 9), 6 (n = 7), 7 (n = 2), 8 (n = 2), 
      12 (n = 5), 13 (n = 15), 14 (n = 1), 16 (n = 17), 17 (n = 11), 19 (n = 21), 20 (n 
      = 12), and/or 22 (n = 17). None of the imbalances were confirmed by fluorescence 
      in situ hybridization (FISH). Using digoxigenin and biotin-conjugated nucleotides 
      in exemplary cases (n = 5) led to the disappearance of the signal deviations. 
      Repeated CGH experiments using seven different reference DNAs showed remarkable 
      variations in the signal deviations. CONCLUSION: Hybridization differences depend 
      not only on the hapten or fluorochrome-labeled nucleotides used for DNA labeling, 
      but also on the reference DNA chosen. Therefore, close control of CGH experiments 
      is mandatory, and additional techniques such as FISH should be performed to 
      confirm the results obtained by CGH.
CI  - Copyright 2003 S. Karger AG, Basel
FAU - Wilkens, Ludwig
AU  - Wilkens L
AD  - Institute of Cell and Molecular Pathology, Hannover Medical School, Hannover, 
      Germany.
FAU - Tchinda, Joelle
AU  - Tchinda J
FAU - Burkhardt, Dagmar
AU  - Burkhardt D
FAU - Kreipe, Hans Heinrich
AU  - Kreipe HH
LA  - eng
PT  - Journal Article
PL  - Switzerland
TA  - Pathobiology
JT  - Pathobiology : journal of immunopathology, molecular and cellular biology
JID - 9007504
RN  - 0 (DNA Probes)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Adult
MH  - *Artifacts
MH  - Bone Marrow Cells/pathology
MH  - Cells, Cultured
MH  - *Chromosome Aberrations
MH  - DNA/*analysis
MH  - *DNA Probes
MH  - Female
MH  - Humans
MH  - In Situ Hybridization/*methods
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Karyotyping
MH  - Leukemia, Myeloid, Acute/genetics/pathology
MH  - Male
MH  - Middle Aged
MH  - Myelodysplastic Syndromes/genetics/pathology
MH  - Myeloproliferative Disorders/genetics/pathology
MH  - Reproducibility of Results
EDAT- 2003/04/08 05:00
MHDA- 2003/11/06 05:00
CRDT- 2003/04/08 05:00
PHST- 2002/06/11 00:00 [received]
PHST- 2002/09/16 00:00 [accepted]
PHST- 2003/04/08 05:00 [pubmed]
PHST- 2003/11/06 05:00 [medline]
PHST- 2003/04/08 05:00 [entrez]
AID - 69330 [pii]
AID - 10.1159/000069330 [doi]
PST - ppublish
SO  - Pathobiology. 2002-2003;70(4):204-8. doi: 10.1159/000069330.