AB056. Establishing the procedure for detection of gr/gr deletions on the Y chromosome in Vietnamese infertile men
Trâm Bảo Nguyễn1, Ngọc-Anh Thị Nguyễn1, Thảo-Trang Nguyễn Phạm1, Trung Tấn Nguyễn2
Background and objective: About 2-10% cases of both azoospermia or cryptospermia have a micro Y deletion. These deletions mainly locate at AZFa, b, or c which containing several crucial genes for normal spermatogenesis. Among various category of deletions is gr/gr deletions on AZFc. The influence on spermatogenesis mostly depend on characteristics of the Y chromosome which may vary among different ethnicity and geographical locations. In Vietnam, there is no study about gr/gr deletions. (I) To establish an approachable procedure to detect gr/gr deletions on the Y chromosome, which is applicable for hospitals and clinics in Vietnam; (II) to determine the percentage of gr/gr deletions carriers among infertile men (azoospermia and severe oligozoospermia).
Methods: Blood samples were collected for DNA extraction. Based on inclusive and exclusive criteria, 3 fertile and 32 infertile men (azoospermia and cryptozoospermia) were recruited to our study with informed consent. Sequence tagged sites (STSs) và primers were designed by using design software (Ape, PrimerPlex2, Oligoanalyzer IDT) based on the information of NCBI. DNA was extracted in silico from blood sample and applied to Multiplex PCR to detect gr/gr deletions. To ensure the true positive result of gr/gr deletions, MLPA was subjected to perform.
Results and discussion: With our designed primers, an approachable procedure was successfully established to detect gr/gr deletions that resulted at a high level of sensitivity (at 0.5ng DNA sample/reaction) and high level of stability. The percentage of gr/gr-deletion carriers among infertile men was 12.5% (4 positive samples), higher than 10.6% in an Asian population from a current study. All positive samples are confirmed significantly by MLPA technique.
Conclusions: An approachable procedure was successfully established to detect gr/gr deletions that resulted at a high level of sensitivity (at 0.5 ng DNA sample/reaction) and high level of stability. The percentage of gr/gr-deletion carriers among azoospermia/ severe oligozoospermia men was 12.5%.
Keywords: gr/gr deletions; multiplex PCR; infertile men
