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.

Volume 98, Issue 4, Pages 843-848.e1, October 2012


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.

  • OperationMiracle

    Even if SSSA doesn’t prevent the need for surgical retrieval, as the wife of a very hopeful azoospermic husband, I can tell you that simply knowing that there ARE sperm would be a huge relief. The not-knowing is very stressful and painful. In addition, there are lots of men who aren’t willing to undergo the surgery because the success rate isn’t high enough. If SSSA could give them more confidence, they might be willing to move forward with the retrieval.

  • khwang

    Completely agree with the comments below. Fascinating technique that clearly still needs to establish it’s clinical relevance as the real utility will be defining how this changes an infertile couples clinical outcome. Will this technique change the need to proceed onto other techniques of sperm retrieval?

  • NicoGarrido

    Congratulations for this nice work
    you are surpassing ione of the pre-existent limits in the andrology laboratory: the misdiagnosis of a male as azoospermic, then referred to TESE, when still has tiny amounts of sperm that were not observed under the standard search
    I hope this technique may be extended in the future and patients showing these extremely low quality samples can be benefited by avoiding unnecesary surgical procedures

  • Pravin Rao

    The comments below raise the most relevant questions regarding this interesting article. However, I also found it Interesting that the authors suggest performing SSSA to increase confidence in vasectomy success. Given that almost half of post-vasectomy samples in some studies have shown low levels of sperm for extended time periods without clinical significance, I fear SSSA would cause unnecessary stress rather than added reassurance in this population.

  • eykko

    It will be interesting to see how this technology evolves. Based on the current processing techniques described in this article, it does not appear that this method of identifying rare sperm would allow the andrologist and embryologist to then retrieve viable sperm for a fresh cycle of ICSI or single sperm vitrification for future attempts. Hopefully, this technology could be refined and ultimately viable sperm could be utilized for ICSI and save a male from undergoing surgical exploration and extration to retrieve sperm.

  • Owning to the high prevalence of azoospermia and the growing recognition of the male contribution to infertility, the work presented by Beyer et al is both timely and significant. As clinicians however, we must be careful how we use the information from SSSA to counsel our patients. The real question is: does the finding of rare sperm change the clinical plan for a given couple? In many cases, the few sperm that are identified in cases of “virtual azoospermia” are not adequate to allow success of a given IVF-ICSI cycle, and surgical sperm retrieval must attempted. In past studies, the finding of cryptozoospermia has failed to predict the success of surgical sperm retrieval. It will be important to determine if SSSA performs better in this capacity or if it changes the final clinical outcome of interest – pregnancy.

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