title: A novel, rapid, and accurate method for detecting microdeletion involving the DAZ gene in infertile men.
abstract: OBJECTIVE
  To report on a novel, accurate method for detecting microdeletion involving the DAZ gene in infertile men.
  
  
  METHODS
  Retrospective clinical study.
  
  
  METHODS
  University Infertility Center of Cochin Hospital, Paris, France.
  
  
  METHODS
  Infertile patients (n = 25) consulting our infertility department during 1998. The patient cohort included subjects with nonobstructive azoospermia and oligoasthenospermia.
  
  
  METHODS
  Blood samples were collected from each subject.
  
  
  METHODS
  DNA analysis using polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE).
  
  
  RESULTS
  We used a new molecular genetic strategy to rapidly identify deletions of the Y chromosome that include the DAZ locus. The experiment consists of amplifying simultaneously exon 4 of the DAZ and DAZLA genes with the use of specific primers that are complementary to intronic sequences of these genes. DGGE was used to separate the two PCR products, with good resolution. In infertile men with a microdeletion of the DAZ gene, this method allows amplification of an internal control when a deletion of that portion of the Yq chromosome is observed on a single amplification.
  
  
  CONCLUSIONS
  This PCR-DGGE method for detection of DAZ gene deletion is simple and fast and does not require the use of radioactive elements. Compared with the classic PCR approach, this new method allows the amplification of the DAZLA copy to be used as an effective internal control in infertile men with microdeletion of the DAZ locus. This procedure could be particularly useful in screening for the DAZ locus in the diagnostic workup of nonobstructive azoospermia and severe oligoasthenoteratozoospermia.
authors.affiliation: Service d'Histo-Embryologie, Biologie de la Reproduction, Hôpital Cochin, Paris, France.
authors.forename: H
authors.initials: H
authors.lastname: Lucas
authors.forename: C
authors.initials: C
authors.lastname: Patrat
authors.forename: P
authors.initials: P
authors.lastname: Jouannet
authors.forename: C
authors.initials: C
authors.lastname: Beldjord
authors.forename: T
authors.initials: T
authors.lastname: Bienvenu
mesh_terms: D000328:Adult; D001483:Base Sequence; D016022:Case-Control Studies; D004252:DNA Mutational Analysis / Q000379:methods*; D000075364:Deleted in Azoospermia 1 Protein; D004586:Electrophoresis / Q000379:methods; D005260:Female; D006801:Humans; D007248:Infertility, Male / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D011506:Proteins / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics*; D012189:Retrospective Studies; D017384:Sequence Deletion; D014998:Y Chromosome
chemical_list: C095668:DAZ1 protein, human; C101358:DAZL protein, human; D000075364:Deleted in Azoospermia 1 Protein; D011506:Proteins; D016601:RNA-Binding Proteins