title: Quantitative fluorescent-PCR detection of sex chromosome aneuploidies and AZF deletions/duplications.
abstract: The most common genetic causes of spermatogenic failure are sex chromosomal abnormalities (most frequently Klinefelter's syndrome) and deletions of the azoospermia factor (AZF) regions (AZFa, AZFb, and AZFc) of the Y chromosome. Several studies have proposed that partial AZFc deletions/duplications may be a risk factor for spermatogenic impairment. We describe a multiplex quantitative fluorescent-polymerase chain reaction (QF-PCR) method that allows simultaneous detection of these genetic causes and risk factors of male infertility. The 11-plex QF-PCR permitted the amplification of the amelogenin gene, four polymorphic X-specific short tandem repeat (STR) markers (XHPRT, DXS6803, DXS981, and exon 1 of the androgen receptor gene), nonpolymorphic Y-specific marker (SRY gene), polymorphic Y-specific STR marker (DYS448), and coamplification of DAZ/DAZL, MYPT2Y/MYPT2, and two CDY2/CDY1 fragments that allow for determination of the DAZ, MYPT2Y, and CDY gene copy number. A total of 357 DNA samples from infertile/subfertile men (n = 205) and fertile controls (n = 152) was studied. We detected 14 infertile males with sex chromosome aneuploidy (10 with Klinefelter's syndrome, 2 XX, and 2 XYY males). All previously detected AZF deletions, that is, AZFc (n8), AZFb (n1), AZFb + c (n1), gr/gr (n11), gr/gr with b2/b4 duplication (n3), and b2/b3 (n5), gave a specific pattern with the 11-plex QF-PCR. In addition, 32 DNA samples showed a pattern consistent with presence of gr/gr or b2/b4 and 4 with b2/b3 duplication. We conclude that multiplex QF-PCR is a rapid, simple, reliable, and inexpensive method that can be used as a first-step genetic analysis in infertile/subfertile patients.
authors.affiliation: Macedonian Academy of Sciences and Arts, Research Center for Genetic Engineering and Biotechnology, Skopje, Republic of Macedonia.
authors.forename: Toso
authors.initials: T
authors.lastname: Plaseski
authors.forename: Predrag
authors.initials: P
authors.lastname: Noveski
authors.forename: Svetlana
authors.initials: S
authors.lastname: Trivodalieva
authors.forename: Georgi D
authors.initials: GD
authors.lastname: Efremov
authors.forename: Dijana
authors.initials: D
authors.lastname: Plaseska-Karanfilska
mesh_terms: D000782:Aneuploidy; D001483:Base Sequence; D016022:Case-Control Studies; D041321:Chromosomes, Human, X / Q000235:genetics; D041322:Chromosomes, Human, Y / Q000235:genetics; D004252:DNA Mutational Analysis; D017931:DNA Primers / Q000235:genetics; D017353:Gene Deletion; D020440:Gene Duplication; D056426:Genetic Loci; D006801:Humans; D007248:Infertility, Male / Q000235:genetics*; D007713:Klinefelter Syndrome / Q000235:genetics; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D029589:Seminal Plasma Proteins / Q000235:genetics*; D012729:Sex Chromosome Aberrations; D025064:Sex Chromosome Disorders / Q000235:genetics; D014997:XYY Karyotype / Q000235:genetics
chemical_list: D017931:DNA Primers; D029589:Seminal Plasma Proteins