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The Relationship between the Halosperm Assay and Semen Analysis Performed According to the 4th and the 5th Editions of the World Health Organization Guidelines

Ashleigh McEvoy, Peter Roberts, Kailin Yap and Phillip Matson

Background: As a standard reference to evaluate male factor infertility, the majority of fertility laboratories use the 4th or 5th Editions of the World Health Organization’s semen analysis guidelines. Following the release of the 5th Edition, debate over its legitimacy has resulted in some laboratories using the 4th and others the 5th Edition. DNA integrity tests have been shown to be a valuable adjunct to semen analysis and have subsequently been adopted by many fertility laboratories. This study explored the prevalence of samples with high DNA fragmentation levels according to semen analysis categories using both the 4th and the 5th Edition reference ranges.

Materials and Methods: The study included 905 consecutive semen samples from 863 infertile couples attending a fertility clinic. A semen analysis was conducted according to both the 4th and 5th Edition guidelines published by the World Health Organization. DNA damage was assessed using the Halosperm G2 test kit and expressed as a percentage DNA fragmentation level.

Results: Alongside both the World Health Organization 4th and 5th Edition semen analysis criteria abnormal DNA fragmentation levels were more common in abnormal semen samples however elevated DNA fragmentation levels were also found in normal semen samples using the same criteria. Of the samples that were graded as normozoospermic according to the 5th Edition guidelines 16% were deemed to have elevated DNA fragmentation levels compared to 11.7% graded by the 4th Edition guidelines. The number of normozoospermic samples, graded according to the 5th Edition guidelines was significantly higher (n=697) than when the same samples were graded according to 4th Edition guidelines (n=385) (p=0.001). A significant proportion of samples with an abnormal DNA fragmentation level corresponding to the World Health Organisation 4th and 5th Edition criteria were evident in normozoospermic (p <0.05), normoteratozoospermic (p=<0.005) and normoasthenozoospermic (p<0.05) samples.

Conclusion: Our findings indicate that abnormal DNA fragmentation levels are proportionate to the World Health Organisation semen analysis criteria with fragmentation levels increasing according to the increasing number of semen analysis abnormalities. In some cases however, abnormal fragmentation levels were recorded when semen analysis was normal and normal fragmentation levels were recorded where the semen analysis was considered abnormal.