Diagnostic yield of OCT3 /4 reflex testing in testicular biopsy

Background and Objective Occult germ cell neoplasia in situ (GCNIS) can occur in testicular biopsies prompted by male infertility with or without concomitant contralateral tumour. Immunohistochemistry can improve the diagnostic yield of GCNIS. The objective of this long‐spanned retrospective single institutional series is to assess the value of OCT3/4 reflex testing on the detection of GCNIS. Methods Through a search of the pathology medical system, we included all men who underwent testicular biopsy for infertility, including cases with a known or suspected germ cell tumour (GCT) between 2010 and 2025. The detection of GCNIS was performed through histology and reflex use of OCT3/4 immunohistochemistry for all cases. Results The detection frequency of GCNIS was 48/3353 (1.4%) testicular biopsies from 42/1870 (2.2%) patients. Subtle interstitial seminoma was diagnosed in 3 patients adjacent to GCNIS. Patients with a concern for GCT were 6x more likely to be diagnosed with GCNIS on biopsy. Among patients with concern for GCT, no difference in GCNIS rate was found between patients with unilateral or bilateral sampling ( P = 0.600). GCNIS was sometimes found adjacent to and even within tubuli with normal spermatogenesis. The frequency of GCNIS was similar ( P = 0.277) among biopsies with no spermatogenesis, diminished spermatogenesis and intact spermatogenesis. Conclusions This study assessed the diagnostic yield of OCT3/4 reflex testing in testicular infertility biopsies with a GCNIS rate of 1.4% and interstitial seminoma frequency of 0.2%. While a history of GCT or clinical suspicion of GCNIS was associated with a 6x higher risk of finding GCNIS, the level of spermatogenesis was not.