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Establishment of a Multiplex – PCR protocol for detection of Y chromosome microdeletion in Azoospermia male patients

Phuong Thi Kim Huynh 1, *
Thanh Kieu Huynh 1
Nam Tri Vo 1
Anh Le Tuan Nguyen 1
Hoang Duc Nguyen 1
Trang Thi Phuong Phan 1
Thong Van Nguyen 2
Giang Ha Pham 2
  1. VNUHCM-University of Science
  2. Hung Vuong Hospital
Correspondence to: Phuong Thi Kim Huynh, VNUHCM-University of Science. Email: pvphuc@vnuhcm.edu.vn.
Volume & Issue: Vol. 18 No. 4 (2015) | Page No.: 5-15 | DOI: 10.32508/stdj.v18i4.904
Published: 2015-12-30

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Copyright The Author(s) 2023. This article is published with open access by Vietnam National University, Ho Chi Minh city, Vietnam. This article is distributed under the terms of the Creative Commons Attribution License (CC-BY 4.0) which permits any use, distribution, and reproduction in any medium, provided the original author(s) and the source are credited. 

Abstract

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.

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