M N Collinson, S Huang, JN Macpherson, VK Maloney, AC Skinner, A Karcanias, NA Affara, NS Thomas
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Transcript of M N Collinson, S Huang, JN Macpherson, VK Maloney, AC Skinner, A Karcanias, NA Affara, NS Thomas
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Discrepant cytogenetic, MLPA and array CGH results on a der(X)(pter->q27.2::p22.31->pter): the
advantages of an integrated approach.
• M N Collinson, S Huang, JN Macpherson, VK Maloney, AC Skinner, A Karcanias, NA Affara, NS Thomas
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30 year old female with premature ovarian failure (POF)
cytogenetic analysis molecular genetic testing for a FRAXA premutation
46,XX only one allele found?45,X or 46,XY
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MLPA with the Rett syndrome kit
MRC-Holland kit PO15C
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LIST OF PROBES USED FOR FISH TO CONFIRM DELETIONCentromere
RP1-142F18 Normal (Xq27.2)
RP1-315J21 Deleted (Xq27.3)
RP11-79A21 Deleted (Xq27.3)
RP11-37P24 Deleted (fragile X locus)
RP11-161L9 Deleted (fragile X locus)
RP11-64L19 Deleted (Xq28)
RP11-533H4 Deleted (Xq28)
RP11-76K17 Deleted (Xq28)
RP11-402H20 Deleted (Xq28)
Telomere
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FISH with a probe for the FRAXA gene
RP11-161L9X centromere (control)
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Analyst Slide
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1 X
2 X
3 X
Blind cytogenetic analysis – 3 control slides and 3 slides from POF patient scored by 3 independent analysts
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Partial karyotype of the X chromosomes(abnormal X on the right)
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MLPA with the subtelomere probe kit
MRC-Holland kit PO36B
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CY29 (Xp subtelomere)
c8.1/2 (Xq subtelomere)
FISH with the Xp and Xq subtelomere probes
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Agilent 244K oligoarray
Array-CGH
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RP11-946P8
DXZ2 (X centromere control)
FISH with SHOX probe
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MLPA with SHOX kit PO18BDuplication of SHOX
POF case
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FEMALE CONTROL MALE CONTROL
Array CGH with chip specifically designed for sex chromosome abnormalities
NORMAL CASE
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FEMALE CONTROL MALE CONTROL
Array CGH with chip specifically designed for sex chromosome abnormalities
POF CASE
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L.M. Russell et al, Cytogenet Genome Res 116:181-185 (2007)
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Original cytogenetic analysis – 1/30 metaphases 45,X
Blind FISH study with X centromere probe (DXZ2)
Metaphases
Interphase cells
POF case Control
1/30 45,X (3.3%) 1/30 45,X (3.3%)
8/100 45,X (8%) 6/100 45,X (6%)
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Summary• An integrated approach was required to establish the
true nature of the X chromosome abnormality
• Highlights the fact that one should be cautious of any shifts from the baseline when analysing array CGH as it may mask significant changes in log ratios
• We still do not have a satisfactory explanation of our results other than mosaicism
• Deletions of this region of Xq have been reported in association with POF