title: A new approach for screening for Y microdeletions: capillary electrophoresis combined with fluorescent multiplex PCR.
abstract: OBJECTIVE
  To apply capillary electrophoresis for rapid screening for Y microdeletions
  
  
  METHODS
  A set of 25 specific sequence tagged sites that cover the azoospermia factor a, b, and c regions of the Y chromosome was amplified in 5 fluorescent multiplex sets each including 5 primer pairs. One of the primers of each pair was labeled with a fluorescent tag attached to the 5'-end. After PCR amplification, analysis of the obtained PCR products was performed using capillary electrophoresis (ABI Prism 3100 Genetic Analyzer). The method was employed to determine Y microdeletions in azoospermic (n = 49) and severe oligozoospermic (n = 149) men.
  
  
  RESULTS
  The number of PCR cycle (from 45 to 30) and the amount of DNA template (20-fold) used in fluorescent multiplex PCR were reduced because of the high sensitivity of capillary electrophoresis. Approximately 1000 multiplex PCR sets from 198 patients were analyzed simultaneously within 50 h. Y microdeletions were found in 3 out of the 198 azoospermic or severe oligozoospermic men.
  
  
  CONCLUSIONS
  Application of capillary electrophoresis for detection of PCR products provides a semiautomated, high throughput method for rapid screening for microdeletions on the Y chromosome.
authors.affiliation: Department of Gynecology, Fertility Clinic, Aarhus University Hospital, Denmark. prbor@hotmail.com
authors.forename: Pinar
authors.initials: P
authors.lastname: Bor
authors.forename: Johnny
authors.initials: J
authors.lastname: Hindkjaer
authors.forename: Steen
authors.initials: S
authors.lastname: Kølvraa
authors.forename: Hans Jakob
authors.initials: HJ
authors.lastname: Ingerslev
mesh_terms: D002872:Chromosome Deletion; D041322:Chromosomes, Human, Y / Q000235:genetics*; D019075:Electrophoresis, Capillary; D006801:Humans; D007248:Infertility, Male / Q000175:diagnosis*; D008297:Male; D009845:Oligospermia / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*