Supersensitive fluorescent semen analysis: Validation on azoospermic and oligozoospermic samples

The improved sensitivity of a fluorescent-based semen analysis technique indicates that a substantial proportion of patients diagnosed as azoospermic by conventional semen analysis may actually have sperm present in their ejaculate.


Claire Elizabeth Beyer, B.Sc., Bettina Kayler, B.Ms. Elissa Osborne, Ph.D., Robert McLachlan, M.D., Tiki Osianlis, Ph.D.

Vol 98, Issue 4, Pages 843-848.e1



To compare the clinical utility of a super sensitive fluorescent semen analysis (SSSA) procedure published by Cooper et al. (2006) with a conventional World Health Organization (WHO) based semen analysis technique in males with severe oligozoospermia or azoospermia who are undergoing fertility assessment.


Prospective single centre study.


IVF clinic.


Patients attending an infertility clinic for semen analysis.



Main outcome measures:

Presence of spermatozoa in the ejaculate.


Semen samples from 100 men were analysed using conventional WHO 4th Edition semen analysis and determined to be either severely oligozoospermic or azoospermic (reported lower limit of detection of 0.1 million sperm/ml). An aliquot of the same unprocessed sample was also analysed using the SSSA protocol (reported lower limit of detection of approximately 8000 sperm/ml). SSSA confirmed the results of conventional semen analysis in 77% of cases. In 22% of cases, sperm were only identified using SSSA. Overall, SSSA was capable of identifying the presence of sperm in significantly more samples than conventional semen analysis.


The reliable differentiation of extreme oligospermia from azoospermia has profound implications in fertility management. This paper provides the first data comparing sperm detection rates using SSSA or conventional WHO-based approaches in extreme oligozoo-/azoospermic men in an IVF setting. Results indicate that approximately 1 in 4 men classified as azoospermic by conventional semen analysis may actually have sperm present. The improved sensitivity of the SSSA technique may be of significant benefit to patients, particularly in fertility and ART decision-making.

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