Sperm penetration assay and its correlation with semen analysis parameters

Laxmi Kant Pandey, Sulekha Pandey, Shraddha Karkare, Ruchi Sinha, Anil K. Gulati


Background: Aim of current study was to determine whether the Sperm Penetration Assay (SPA) can be used as a test to discriminate the infertile male from fertile one. We have also correlated the SPA with semen analysis.

Methods: Sperm characteristics namely Semen analysis and the sperm penetration assay were tested in 44 infertile and 10 fertile men. Sperm penetration assay was determined by using zona free hamster eggs.

Results: With decreasing spermatozoa concentration in the semen there was significant decrease in percentage penetration of zona free Hamster eggs (p<0.001). There was decrease in Sperm penetration assay with deteriorating progressive sperm motility (p<0.05) and no consistent relationship appeared between the sperm morphology and the Sperm penetration assay (p>0.05).  

Conclusions: The Sperm penetration assay could discriminate the infertile group from fertile group significantly (p<0.001). The test appeared to be highly reproducible and probably identifies a truly infertile male.  


Sperm penetration assay (SPA), Semen analysis, Infertility, Zona free, Hamster oocyte

Full Text:



Guzick DS, Overstreet JW, Factor-Litvak P, Brazil CK, Nakajima ST, Coutifaris C, et al. Sperm morphology, motility, and concentration in fertile and infertile men. N Eng J Med. 2001;345:1388-93.

Cooper TG, Noonan E, von Eckardstein S, Auger J, Baker HW, Behre HM, et al. World Health Organization reference values for human semen characteristics. Hum Reprod Update. 2010;16:231-45.

Shibahara H, Mitsuo M, Inoue M, Hasegawa A, Shigeta M, Koyama K. Relationship between human in-vitro fertilization and intracytoplasmic sperm injection and the zona-free hamster egg penetration test. Hum Reprod. 1998;13:1928-32.

Carrell DT. Semen analysis at the turn of the century: an evaluation of potential uses of new sperm function assays. Arch Androl. 2000;44:65-75.

Blasco L. Clinical test of sperm fertilizing ability. Fertil Steril. 1984;41(2):177.

Yanagimachi R, Yanagimachi H, Rogers BJ. The use of zona-free animal ova as a test-system for the assessment of the fertilizing capacity of human spermatozoa. Biol Reprod. 1976;15:471-6.

Rogers BJ, Van Campen H, Ueno M, Lambert H, Bronson R, Hale R. Analysis of human spermatozoal fertilizing ability using zona-free ova. Fertil Steril. 1979;32:664-70.

Soffer Y, Golan A, Herman A, Pansky M, Caspi E, Raphael RE. Prediction of in vitro fertilization outcome by sperm penetration assay. Fertil Steril. 1992;58:547-50.

Rogers B J. The sperm penetration assay - Its usefulness re-evaluated. Fertil Steril. 1985;43(6):821-40.

Corson SL. Value of hamster egg penetration assay. Am J Obstet Gynecol. 1990;162(3):865.

Rashid MRZ, Fishel SB, Thornton S, Hall JA, Ndukwe G, Aloum M, et al. The predictive value of the zona-free hamster egg penetration test in relation to in-vitro fertilization at various insemination concentrations. Hum Reprod. 1998;13:624-9.

O’Shea DL, Odem RR, Cholewa C, Gast MJ. Long-term follow-up of couples after hamster egg penetration testing. Fertil Steril. 1993;60:1040-5.

Stenchever MA, Spadoni LR, Smith WD, Karp LE, Shy KK, Moore DE, et al. Benefits of the sperm (hamster ova) penetration assay in the evaluation of the infertile couple. Am J Obstet Gynecol. 1982;143:91.

Freeman MR, Archibong AE, Mrotek JJ, Whitworth CM, Weitzman GA, Hill GA. Male partner screening before in vitro fertilization: preselecting patients who require intracytoplasmic sperm injection with the sperm penetration assay. Fertil Steril. 2001;76:1113-8.

World Health Organization. World Health Organization laboratory manual for the examination of human Semen and Semen-cervical mucus interaction. Cambridge, UK: Cambridge University Press; 1999.

Chard T, Lilford RJ. How useful is a test? In: John Studd, eds. Progress in Obstetrics and Gynecology. 9th ed. London: Churchill Livingstone; 1991.

Martin RH, Taylor PJ. Reliability and accuracy of the zona-free hamster ova assay in assessment of male fertility. Br J Obstet Gynaecol. 1982;89:951-6.

Karp LE, Williamson RA, Moore DE, Shy KK, Plymate SR, Smith WD. Sperm penetration assay: useful test in evaluation of male fertility. Obstet Gynecol. 1981;57:620-3.

Overstreet JW, Hembree WC. Penetration of the zona pellucid of non-living human oocyte by human spermatozoa in vitro. Fertil Steril. 1980;33:534.