Screening of subfertile men for testicular carcinoma in situ by an automated image analysis‐based cytological test of the ejaculate

Abstract

Testicular cancer (TC) is usually diagnosed after manifestation of an overt tumour. Tumour formation is preceded by a pre-invasive and asymptomatic stage, carcinoma in situ (CIS) testis, except for very rare subtypes. The CIS cells are located within seminiferous tubules but can be exfoliated and detected in ejaculates with specific CIS markers. We have built a high throughput framework involving automated immunocytochemical staining, scanning microscopy and in silico image analysis allowing automated detection and grading of CIS-like stained objects in semen samples. In this study, 1175 ejaculates from 765 subfertile men were tested using this framework. In 5/765 (0.65%) cases, CIS-like cells were identified in the ejaculate. Three of these had bilateral testicular biopsies performed and CIS was histologically confirmed in two. In total, 63 bilateral testicular biopsy were performed in conjunction with analysis of the ejaculates because of infertility work-up. Histological analysis of the biopsies for the presence of CIS yielded a test sensitivity of 0.67 and a specificity of 0.98. In addition, ejaculates from 45 patients with clinical signs of an overt TC were investigated and yielded a slightly lower sensitivity (0.51), possibly because of obstruction. We conclude that this novel non-invasive test combining automated immunocytochemistry and advanced image analysis allows identification of TC at the CIS stage with a high specificity, but a negative test does not completely exclude CIS. On the basis of the results, we propose that the assay could be offered to subfertile men and other patients who are at increased risk of TC.

The genomic landscape of testicular germ cell tumours: from susceptibility to treatment

“…The invasive nature of testicular biopsy currently restricts development of TGCT screening programmes. Improvements in sensitivity and specificity of semen assay, an emer ging noninvasive diagnostic tool by which presence or absence of ITGCN is detected via staining for fetal germ cell markers, could influence the feasibility of TGCT screening in the future 118,119 . Such an approach would be taking advantage of the unique characteristic of ITGCN as a faithful precursor lesion of TGCT, clinically detectable from adolescence.…”

Diagnostic markers for germ cell neoplasms: from placental-like alkaline phosphatase to micro-RNAs

“…Subsequently, our group has improved sensitivity of the AP-2g-based test by combining it with rapid histochemical staining detecting enzymatic activity of PLAP [82]. The double-staining method was further optimised by automated staining, slide scanning and imaging software, which increased the speed and objectivity of the test [83]. An example of GCNIS cells detected by this method is shown as an insert in Figure 1.…”

Testicular Dysgenesis Syndrome, Cryptorchidism, Hypospadias, and Testicular Tumors

“…Because of the special consistency of the testis tissue, it is recommended that the biopsy adheres to certain technical requirements. In recent years, progress has been made toward establishing a noninvasive test to detect CIS cells in semen by immunocytochemistry, [184][185][186] and this approach may in the future eliminate the need for the contralateral biopsy. Therefore, many European centers advocate performing a contralateral biopsy simultaneously with an orchidectomy procedure.…”