A rapid and simple system of detecting deletions on the Y chromosome related with male infertility using multiplex PCR.
Around 10% of males with idiopathic azoospermia or oligozoospermia, which are important causes of male infertility, have partial deletions on the long arm of the Y chromosome. To develop a rapid and accurate detection system for screening major Y deletions found in infertile men, we developed a multiplex PCR system that can simultaneously amplify five loci on the Y chromosome, SRY, AMELY, DBY, RBMY, DAZ and one locus on the X chromosome, AMELX. The size of the PCR products was designed to increase gradually from the distal Yp to the distal Yq. Our system could detect deletions of three major candidate regions for the azoospermic factor, AZFa, AZFb and AZFc on the Y chromosome together with sex. The gradual increase in the size of the PCR products was convenient for imaging the location of deletions on the Y chromosome. Moreover, the multiplex PCR system was combined with microchip-based electrophoresis, and the PCR products derived from each locus were separated within 4 min. Our system is useful for screening Y chromosomes bearing the structural anomalies including three major AZF deletions found among azoospermic or oligozoospermic males.
Umeno
Mayumi
M
Department of Human Genetics and Public Health, Institute of Health Biosciences, The University of Tokushima Graduate School, Japan.
Shinka
Toshikatsu
T
Sato
Youichi
Y
Yang
Xin-Jun
XJ
Baba
Yoshinobu
Y
Iwamoto
Teruaki
T
Nakahori
Yutaka
Y
D001483:Base Sequence; D016022:Case-Control Studies; D002872:Chromosome Deletion; D041322:Chromosomes, Human, Y / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004586:Electrophoresis / Q000379:methods; D056426:Genetic Loci; D006801:Humans; D008297:Male; D009845:Oligospermia / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D029589:Seminal Plasma Proteins / Q000235:genetics
D017931:DNA Primers; D029589:Seminal Plasma Proteins