Main Article Content


A.D. Kairzhanova

National Center for Biotechnology, 13/5, Korgalzhyn road, Astana, 010000, Kazakhstan

G.D. Abisheva

National Center for Biotechnology, 13/5, Korgalzhyn road, Astana, 010000, Kazakhstan

O.A. Popova

Astana Ecolife, 22, Alash road, Astana, 010000, Kazakhstan

D.K. Kamalova

National Center for Biotechnology, 13/5, Korgalzhyn road, Astana, 010000, Kazakhstan

E.S. Shevtsova

National Center for Biotechnology, 13/5, Korgalzhyn road, Astana, 010000, Kazakhstan

A.B. Shevtsov

National Center for Biotechnology, 13/5, Korgalzhyn road, Astana, 010000, Kazakhstan


Microdeletions in the long arm of the Y chromosome are common causes of male spermatogenesis disorders. The frequency of the AZF locus microdeletions is approximately1 in 1000–1500 males. Such deletions of the Y chromosome are identified in 11% of males with azoospermia and in 8% of males with severe oligospermia. The aim of this study was to develop a multiplex polymerase chain reaction (PCR) protocol for the detection of microdeletions in the AZF locus of the Y chromosome. Based on the literature, the following sequence tagged site (STS) markers were used for development of the PCR protocol for screening the AZF locus microdeletions: AZFa-sY86 иsY84, AZFb-sY127 and sY134, AZFс-sY254 and sY255. As a control, the presence of Y chromosome short arm fragments in the genomic DNA were assessed using the SRY gene sequence and ZFY/Х as an internal control of the PCR. Confirmation of the developed protocol was performed with 40 DNA samples. The protocol recommended by The European Academy of Andrology was used as a complementary method to confirm our results. The results from both methods indicate the specificity and high demand of the developed PCR protocol.


male infertility, PCR, microdeletion, AZF locus, Y chromosome, STS markers

Article Details


Esteves S.C. A clinical appraisal of the genetic basis in unexplained male infertility. J. Hum Reprod Sci, 2013, vol. 6, pp. 176-82. doi: 10.4103/0974-1208.121419.

Rowe P.J., Comhaire F.H. WHO manual for the standardised investigation and diagnosis of the infertile couple. Cambridge: Cambridge University, 2000.

Farhi J., Ben-Haroush A. Distribution of causes of infertility in patients attending primary fertility clinics in Israel Isr. Med. Assoc. J., 2011, vol. 13, no 1, pp. 51-54. PMID: 21446238.

Nieschlag E., Behre H.M., Nieschlag S. Andrology: male reproductive health and dysfunction. Springer; Heidelberg; New York, 2010, pp. 577-587.

Chernih V.B., Kurilo L.F., Gogolevskaya I.K. et. al. Complecsnoe clinic-geneticheskoe bsledovanei pacientov c azoospermiey ili oligozoospermiey neyasnoi etiologii [Complex clinical-genetic examination of patients with azoospermia or oligozoospermia unknown etiology]. Problemi reprodukcii – Reproduction problems, 2001, no 3, pp. 58-63.

Scriven P.N., Flinter F.A., Braude P.R., Ogivie C.M. Robertsonian translocations-reproductive risks and indications for preimplantation genetic diagnosis. Hum.Reprod, 2001, vol. 16, pp. 2267-2273. PMID:11679502.

Reynolds N., Cooke H.J. Role of the DAZ genes in male fertility. Reprod Biomed Online, 2005, vol. 10, pp. 72-80. PMID: 15705297.

Foresta C., Moro E., Ferlin A. Y chromosome microdeletions and alterations of spermatogenesis. Endocrine Reviews, 2001, vol. 22, pp. 226-329. PMID:11294825.

Krausz C., Hoefsloot L., Simoni M., Tüttelmann F. EAA/EMQN best practice guidelines for molecular diagnosis of Y-chromosomal microdeletions: state-of-the-art 2013. Andrology, 2014, vol. 2, pp. 5-19. doi: 10.1111/j.2047-2927.2013.00173.x.

Simoni M., Tüttelmann F., Gromoll J., Nieschlag E. Clinical consequences of microdeletions of the Y chromosome: the extended Münster experience. Reprod Biomed Online, 2008, vol. 16, pp. 289-303. PMID:18284889.

Vogt P.H. Azoospermia factor (AZF) in Yq11: towards a molecular understanding of its function for human male fertility and spermatogenesis. Reprod. Biomed. Online, 2005, vol. 10, pp. 81-93. PMID: 15705299.

Navarro-Costa P., Plancha C., Gonçalves J. Genetic dissection of the AZF regions of the human Y chromosome: thriller or filler for male (in)fertility? Journal of Biomedicine and Biotechnology, 2010, vol. 16, pp. 525-542. PMID: 20671934.

Krausz C. Y chromosome and male infertility: update. Degl. Innocenti. S.Front. Biosci, 2006, vol. 11, pp. 3049-3061. PMID:16720375.

Ferlin A., Raicu F., Gatta V., Zuccarello D., Palka G., Foresta C. Male infertility: roleofgenetic background. Reprod. Biomed. Onlin, 2007, vol. 14, pp. 734-45. PMID: 17579990.

Nuti F., Krausz C. Gene polymorphisms/mutations relevant to abnormal spermatogenesis. Reprod. Biomed. Online, 2008, vol. 16, pp. 504-13. PMID: 18413059.

Brown G.M., Furlong R.A., Sargent C.A., Erickson R.P., Longepied G., Mitchell M. et al. Characterisation of the coding sequence and fine mapping of the human DFFRY gene and comparative expression analysis and mapping to the Sxrb interval of the mouse Y chromosome of the Dffry gene. Hum. Mol. Genet, 1998, no 7, pp. 97-107. PMID: 9384609.

Ferlin A., Arredi B., Speltra E., Cazzadore C., Selice R., Garolla A. et al. Molecular and clinical characterization of Y chromosome microdeletions in infertile men: a 10-year experience in Italy. Journal Clin. Endocrinol. Metab, 2007, vol. 92, pp. 762-70. PMID: 17213277.

Ferlin A., Moro E., Garolla A., Foresta C. Human male infertility and Y chromosome deletions: role of the AZF-candidate genes DAZ, RBM and DFFRY. Hum. Reprod, 1999, vol. 14, pp. 1710-1716. PMID:10402373.

Georgiou I., Syrrou M., Pardalidis N., Karakitsios K., Mantzavinos T., Giotitsas N. et al. Genetic and epigenetic risks of intracytoplasmicsperm injection method. Asian. J. Androl, 2006, vol. 8, pp. 643-673. doi: 10.1111/j.1745-7262.2006.00231.x.

Hellani A., S.Al-Hassan et al. Y chromosome microdeletions: are they implicated in teratozoospermia? Hum. Reprod, 2005, vol. 2, pp. 3505-3509. PMID:16123092.

Sadeghi-Nejad. H. and Farrokhi F. Genetics of azoospermia: current knowledge, clinical implications, and future directions. Part II: Y chromosome microdeletions. Urol J., 2007, vol. 4, no 4, pp. 192-206. PMID:18270942.

Miller S.A., Dykes D., Polesky H.F. A simple salting–out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res, 1988, vol. 16, pp. 1215. doi: 3344216.

Zheng H.Y., Li Y., Shen F.J., Tong Y.Q. A novel universal multiplex PCR improves detection of AZFc Y-chromosome microdeletions. J Assist. Reprod. Genet, 2014, vol. 31, no 5, pp. 613-620. doi: 10.1007/s10815-014-0204-5.

Shevtsov A.B., Kairzhanova A.D., Abisheva G.D. et al. Razrabotka pcr testa dlya vidovoi identificasii C. coli, C. jejuni, C. fetus. [Development pcr test for specific identification c. Coli, c. Jejuni, c. Fetus]. Biotekhnologiya. Teoriya i practika – Biotechnology. Theory and Practice, 2014, no 3, pp. 54-60. doi: 10.11134/

Roux K.H. Cold Optimization and troubleshooting in PCR. Spring Harbor protocols, 2009, pp. 380. doi: 10.1101/pdb.ip66.

Gunson R.N., Bennett S., Maclean A., Carman W.F. Using multiplex real time PCR in order to streamline a routine diagnostic service. J. Clin Virol, 2008, vol. 43, no 4, pp. 372-375. doi:10.1016/j.jcv.2008.08.020.