title: Rapid detection of chromosomes X and Y aneuploidies by quantitative fluorescent PCR.
abstract: Quantitative fluorescent polymerase chain reaction (QF-PCR) assays and small tandem repeat (STR) markers have been successfully employed for the rapid detection of major numerical aneuploidies affecting human autosomes. So far, the analysis of chromosomes X and Y disorders has been hampered by the rarity of highly polymorphic markers which could distinguish normal female homozygous PCR patterns from those seen in patients with Turner's syndrome. A new marker (X22) of the X/Y chromosomes has been identified which maps in the Xq/Yq pseudoautosomal region PAR2; used together with the HPRT it allows the rapid diagnosis of numerical aneuploidies of the sex chromosomes. Blood samples from normal male and female subjects and from patients with X and Y chromosome disorders (45,X and 47, XXY) have been tested by QF-PCR with the X22 polymorphic pentanucleotide (12 alleles) together with the HPRT and P39 markers. The samples were also tested by multiplex QF-PCR with STRs specific for chromosomes 21,18,13 and amelogenin (AMXY). Tested by QF-PCR, all samples from normal females were heterozygous for either the X22 or the HPRT marker with fluorescent peak ratios near 1:1, thus allowing a correct, rapid diagnosis of their chromosome complement. Turner's patients (45,X) showed only one X22 and one HPRT fluorescent peak, thus documenting the presence of a single X chromosome. Turner's patients with mosaicism showed a major fluorescent peak for the X22 and HPRT markers and a minor peak revealing the presence of a second minor population of cells. Two 47, XXY cases could also be diagnosed. Multiplex analyses can be performed using simultaneously STR markers for chromosomes 21,18,13 X and Y. The diagnostic value of a third X-linked marker (P39) was also investigated. These results suggest that rapid diagnosis of major numerical anomalies of the X and Y chromosomes can be performed using QF-PCR with a new highly polymorphic X-linked marker, X22, which maps in the Xq/Yq pseudoautosomal region PAR 2. Multiplex QF-PCR tests-using the X22 STR in association with HPRT and, in rare cases, a third P39 marker-allow the rapid diagnosis of major aneuploidies affecting chromosomes 21, 18, 13, X and Y. The X22 marker can also be employed for the detection of fetal cells present in maternal peripheral blood or the endocervical canal.
authors.affiliation: Galton Laboratory and Department of Obstetric and Gynaecology, University College London, London, U.K.
authors.forename: V
authors.initials: V
authors.lastname: Cirigliano
authors.forename: J
authors.initials: J
authors.lastname: Sherlock
authors.forename: G
authors.initials: G
authors.lastname: Conway
authors.forename: C
authors.initials: C
authors.lastname: Quilter
authors.forename: C
authors.initials: C
authors.lastname: Rodeck
authors.forename: M
authors.initials: M
authors.lastname: Adinolfi
mesh_terms: D000782:Aneuploidy; D016022:Case-Control Studies; D005260:Female; D005453:Fluorescence; D005819:Genetic Markers; D006801:Humans; D007621:Karyotyping; D008297:Male; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D014424:Turner Syndrome / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D014960:X Chromosome; D014998:Y Chromosome
chemical_list: D005819:Genetic Markers