Reverse chromosome painting: a method for the rapid analysis of aberrant chromosomes in clinical cytogenetics.
We describe a method, termed reverse chromosome painting, which allows the rapid analysis of the content and breakpoints of aberrant chromosomes. The method involves the sorting of small numbers of the aberrant chromosome from short term blood culture preparations or cell lines by using bivariate flow karyotype analysis. The sorted chromosomes are amplified and biotin labelled enzymatically using a degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), the product annealed to metaphase spreads from normal subjects, and hybridisation detected using fluorescence in situ hybridisation (FISH). We show the usefulness of this method for routine clinical cytogenetics by the analysis of cases involving an insertion, a deletion, a translocation, and two cases of a chromosome with additional material of unknown origin. The method has particular application for the rapid resolution of the origin of de novo unbalanced chromosome duplications.
Carter
N P
NP
Department of Pathology, University of Cambridge.
Ferguson-Smith
M A
MA
Perryman
M T
MT
Telenius
H
H
Pelmear
A H
AH
Leversha
M A
MA
Glancy
M T
MT
Wood
S L
SL
Cook
K
K
Dyson
H M
HM
D000328:Adult; D001483:Base Sequence; D002478:Cells, Cultured; D002648:Child; D002869:Chromosome Aberrations; D002871:Chromosome Banding; D002872:Chromosome Deletion; D003582:Cytogenetics / Q000379:methods*; D004247:DNA; D015342:DNA Probes; D005069:Evaluation Studies as Topic; D005260:Female; D005434:Flow Cytometry; D005453:Fluorescence; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007621:Karyotyping; D008297:Male; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D014178:Translocation, Genetic
D015342:DNA Probes; D004247:DNA