Detection of microdeletions in the short arm of the X chromosome by chromosome stretching

The present study was focused on the resolution of "chromosome stretching". In order to determine if this method can be used for the detection of microdeletions, the p-arms of 13 normal X chromosomes were stretched as well as of those with three different deletions of known size within the DMD/BMD region in Xp21 (case A: 0.42-0.45 Mb, case B: 2.3-19 Mb and case C: 3.0-3.5 Nib). The process of band splitting was recorded on a video-tape and the resulting banding pattern analyzed. Stretching of the normal Xp-arms led to a splitting on a maximum band level of 1400 and showed in all cases an identical banding pattern with 13 Giemsa-dark subbands. All new Giemsa-dark and -light subbands were derived from the three initial Giemsa-dark bands at the 400 band level according to ISCN (1995): five subbands from Xp21, four subbands from Xp11.3 and Xp22.2, respectively. The origin of these subbands is partly in contrast to the high resolution ISCN (1995) ideograms: subband Xp11.22 does not originate from the Giemsa-light band Xp11.2, but from the Giemsa-dark band Xp11.3; Xp22.12 originates from Xp21; Xp22.32 from Xp22.2. Stretching of the chromosomes containing deletions showed in cases A and B no differences in banding patterns and splitting order compared to normal X chromosomes. Only in patient C was a significant difference with the normal pattern visible due to the absence of one dark subband. In this case only four Giemsa-dark subbands derived from band Xp21. Thus, at least in the DMD/BMD region, the minimal size of a deletion detected by chromosome-stretching-generated high-resolution ideograms is about 3.0-3.5 Mb. Copyright (C) 2002 S. Karger AG, Basel.