Quantification of X-chromosome inactivation patterns in haematological samples using the DNA PCR-based HUMARA assay

作者: C A Mein , D C Linch , R E Gale

DOI:

关键词: X-inactivationAlleleLocus (genetics)Molecular biologyBiologyBlotMyeloidSouthern blotPolymerase chain reactionX chromosome

摘要: Quantification of X-chromosome inactivation patterns (XCIPs) using PCR amplification the human androgen receptor (HUMARA) locus is potentially valuable in a range haematological disorders. Of 236 females screened, 203 (86%) were heterozygous. For quantitative XCIPs it was necessary to limit number cycles 20 reduce preferential shorter alleles. The optimized method compared with Southern blotting results either PGK, HPRT or M27beta 51 haematologically normal and blast cells from 27 patients acute myeloid leukaemia (AML). Reproducible XCIP obtained all 78 samples digestion Hpa II prior (median difference duplicate values 3%, 0-17%) they correlated well results, r=0.966. Greater variability observed Hha I 4%, 0-48%). There marked inconsistencies repeated analyses three AML although HUMARA-Hha overall remaining 75 (r=0.922), nine there still discrepancies > = 20% between two values. These suggest that analysis HUMARA II-digested DNA suitable for quantification but should be treated caution.

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