Open AccessEstablishment of a Multiplex – PCR protocol for detection of Y chromosome microdeletion in Azoospermia male patients
Author(s) -
Phuong Truc Huynh,
Thanh Kieu Huynh,
Nam Tri Vo,
Anh Thu Nguyen,
Hoàng Văn Nguyễn,
Trang Thi Phuong Phan,
Thong Van Nguyen,
Giang Ha Pham
Publication year - 2015
Publication title -
khoa học công nghệ
Language(s) - English
Resource type - Journals
ISSN - 1859-0128
DOI - 10.32508/stdj.v18i4.904
Subject(s) - azoospermia factor , multiplex polymerase chain reaction , y chromosome , testis determining factor , genetics , y chromosome microdeletion , sequence tagged site , azoospermia , biology , multiplex , male infertility , primer (cosmetics) , chromosome , gene , polymerase chain reaction , infertility , karyotype , gene mapping , chemistry , pregnancy , organic chemistry
Microdeletion on the Y chromosome is one of the causes that makes men infertile, accounting for 2-10 % of all infertility cases, and occurs frequently at 3 regions of the Ychromosome long arm namely AZFa, AZFb and AZFc (azoospermia factor). Currently, the diagnosis of microdeletion on the Y chromosome is almost mandatory in institutes and centers for infertility diseases before selecting treatment or assisting methods. To detect microdeletion in AZF, SRY and ZFY regions, the current approach is a Multiplex – PCR assay offering by European Academy of Andrology/European Molecular Genetics Quality Network (EAA/ EMQN). However, the drawback of this method is the PCR products posess similar size and then the DNA electrophoresis bands were very close on gels causing the difficult in diagnosis. Therefore, in this study, we have redesigned primer pairs matching with genes that were recommended by EAA/EMQN but the PCR products are clearly different in sizes, making the DNA electrophoresis bands take apart further to facilitate the diagnosis. Besides, we have also created recombinant plasmids carrying the marker genes for the control sample in kits.