[Using multiplex PCR to analyze the breakpoint of a severe Y-chromosome deletion].
OBJECTIVE
To elucidate the relationship between azoospermia factor(AZF) microdeletion of Y chromosome and azoospermia, the exact breakpoint of a severe Y-chromosome deletion was determined according to the physical map of AZF.


METHODS
Multiplex polymerase chain reaction was used to amplify fifteen sequence tagged sites (STS), namely sY82, sY84, sY86 in AZFa, sY124, sY127, sY128, sY133, sY134, sY143 in AZFb, sY239, sY242 sY254, sY255 in AZFc, and sY145, sY152 in AZFd; sex-determining region Y(SRY) was taken as an internal control. And then sY82,sY86,sY85,sY84 were further analyzed using the sample of the patient who had Y-chromosome deletion by G band analysis to map the breakpoint at molecular level.


RESULTS
All 15 STS and sY85 were amplified in positive control while only sY82, sY86 were amplified in the clinical sample, thus the breakpoint was found to be between sY86 and sY85.


CONCLUSIONS
This study on the patient provided the direct biomolecular evidence of the exact breakpoint of the severe Y-chromosome deletion and established the deletion map of acrocentric chromosome. It also proved that the patient's azoospermia was due to the deletion of AZF.
Xiao
Xiao-su
XS
Laboratory of Genetics, Reproductive Medical Center, Shenzhen Hospital of Beijing University, Shenzhen, Guangdong, PR China. xiaosu-xiao@yahoo.com.cn
Liu
Xiao-yi
XY
Wang
Yong-qiang
YQ
Zhou
Yi
Y
Ouyang
Li
L
Zhang
Xu
X
Cai
Zhi-ming
ZM
D053713:Azoospermia / Q000175:diagnosis / Q000235:genetics*; D019457:Chromosome Breakage; D002872:Chromosome Deletion; D041322:Chromosomes, Human, Y / Q000235:genetics*; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity