Assessment of the Separation of X- and Y-Chromosome Bearing Sperms of Fertile Men by Albumin Gradients Using Real Time PCR
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Date
2016-04-01
0000-0001-5268-8407 Journal Title
Journal ISSN
Volume Title
Publisher
Brieflands
Abstract
Background: One of the techniques used in infertility centers for separating the spermatozoa containing Y or X chromosomes is the Ericsson method. It is simple, cheap, and practical without using any toxic material. There are contradictory reports regarding the efficiency of this extremely reliable method. Objectives: The study aimed to assess the separation of X- and Y-chromosome bearing sperms of fertile men on albumin gradients using real time PCR. Patients and Methods: In this study, 20 samples of human semen were investigated using Ericsson method. After preparation of samples with Ham’s F10 medium, X- and Y-chromosome bearing sperms of samples were separated using albumin gradient, and sperms DNA was extracted by Bioneer kit. Using primers designed for SRY and HPRT genes and ALBUMIN gene, PCR was performed with real time method using SYBR green method, also ΔΔCt method analysis was used to measure the amount of sperms with Y chromosome. Data were analyzed using t test by SPSS and P < 0.05 was considered as significance level. Results: Mean percentage of Y-bearing sperms in the albumin and the control groups were 75.80% (20 samples of human semen) and 49.48%, respectively. Comparing the mean percentages of Y-bearing sperms in albumin and control groups, the difference was found statistically significant (P = 0.001). Conclusions: Albumin gradient method is the enrichment of Y-bearing chromosome sperms. Separation of spermatozoa by albumin gradient method, introduced by Ericsson, is a suitable method for sex selection of male offspring. This technique is safe, because it neither requires toxic materials during sperm separation, nor involves embryo manipulation.