A new approach for screening for Y microdeletions: capillary electrophoresis combined with fluorescent multiplex PCR.
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.
Bor
Pinar
P
Department of Gynecology, Fertility Clinic, Aarhus University Hospital, Denmark. prbor@hotmail.com
Hindkjaer
Johnny
J
Kølvraa
Steen
S
Ingerslev
Hans Jakob
HJ
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*