Research Center for Genetic Engineering and Biotechnology “Georgi D. Efremov”, MASA

What is infertility?Infertility is defined as a failure to conceive in a couple trying to reproduce for a period of one year without conception. Infertility is a major health problem today, affecting about 15% of couples trying to have a child. Impaired fertility of the male factor is causative in 20% of infertile couples and contributory in up to another 30-40%.

What are genetic causes for male infertility?Microdeletions of the azoospermia factor (AZF) regions of the Y chromosome and sex chromosomal aneuploidy (extra or missing chromosome) are the most common known genetic causes of spermatogenic failure. The frequency of these two genetic anomalies increases with the severity of the spermatogenic defect, reaching to an overall 30% (15% karyotype abnormalities and 15% of AZF microdeletions) in azoospermic men. Two additional mutations on autosomal chromosomes with negative effect on male fertility are cystic fibrosis (mutations in CFTR gene) and globozoospermia (deletion of DPY19L2 gene).Who should be tested?Y-chromosome microdeletions and sex chromosome aneuplodies are relatively frequent and their frequency increases with the severity of the spermatogenic defect. Therefore, testing for this conditions is indicated in men with azoospermia (absence of spermatozoids in the semen) and severe oligozoospermia (or sperm concentration <1 x 106/mL), and in patients considering assisted reproduction. Patients with congenital absence of the vas deferens and obstructive azoospermia are strongly indicated for CFTR mutations screening, while patients with presence of round headed spermatozoa in their semen should be tested for DPY19L2 deletion.

Importance of genetic testingThe detection for genetic causes of male infertilty may provide the diagnosis for infertility and allow physician to direct patients to assisted reproduction and to provide proper genetic counseling. Since sons of men with Y-chromosome microdeletions conceived with assisted reproduction will be infertile, child's male factor infertility needs to be discussed. For patients diagnosed with CF or carriers of CFTR mutation, screening for CFTR mutations in female partner should be offered to asses the possibility of having a child with CF. Patients with globozoospermia and Klinefelter syndrome could be advised for proper assisted reproduction technique.

Tests performed at RCGEB Price (МКД)Microdeletions of Y chromosome and sex chromosomes aneuploidy

7200

Screening for CFTR mutations using SNaPshot method 5000

Detection of DPY19L2 gene deletion in globozoospermic men 4600

LiteratureLiterature::

1.Simoni M, Bakker E, Krausz C. EAA/EMQN best practice guidelines for molecular diagnosis of y-chromosomal microdeletions. State of the art 2004. Int J Androl. 2004 Aug;27(4):240-9.2.Plaseski T, Noveski P, Trivodalieva S, Efremov GD, Plaseska-Karanfilska D. Quantitative fluorescent-PCR detection of sex chromosome aneuploidies and AZF deletions/duplications. Genet Test. 2008 Dec;12(4):595-605. doi: 10.1089/gte.2008.0068.3.Jungwirth, A., A. Giwercman, et al. (2012). "European Association of Urology guidelineson Male Infertility: the 2012 update." Eur Urol 62(2): 324-332.4.WHO/Infertility, " http://www.who.int/topics/infertility/en/". Who.int. 2013-03-19. Retrieved 2013-06-17.5.Noveski, P., S. Madjunkova, et al. (2013). "A Homozygous Deletion of the DPY19l2 Gene is a Cause of Globozoospermia in Men from the Republic of Macedonia." Balkan J Med Genet 16(1): 73-76.

Test methods performed at RCGEB “Georgi D. Efremov”Y chromosome microdeletions is currently diagnosed by using polymerase chain reaction amplification (PCR) and gel electrophoresis in order to test whether the DNA sequence corresponding to the selected markers is present in the DNA.Additionally, sex chromosome aneuploidies and AZF deletions/duplications are detected with multiplex quantitative fluorescent-polymerase chain reaction (QF-PCR) with simultaneous amplification of 13 markers.Presence of CFTR mutations is evaluated with single nucleotide primer extension (SNaPshot®) method for analysis of 11 most common CFTR mutations and IVS8 polyT sequence.The deletion of DPY19L2 in globozoospermiac men is evaluated with PCR analysis of seven STS markers (3 in exons and 4 outside of the gene) followed by gel electrophoresis and by MLPA analysis.Material for testingWhole blood specimens in sterile tubes with anticoagulant EDTA

RCGEB, 2013

Genetic testing for male infertilityGenetic testing for male infertility