Unbiased enrichment of urine exfoliated cells on nanostructured substrates for sensitive detection of urothelial tumor cells

Abstract

BackgroundEarly detection of urothelial carcinoma (UC) by noninvasive diagnostic methods with high accuracy is still underscored. This study aimed to develop a noninvasive assay incorporating both enrichment of urine exfoliated cells and immunoassays for UC detection.MethodsPolystyrene dishes were exposed to oxygen plasma and modified with 3‐aminopropyltriethoxysilane to prepare amine‐functionalized nanostructured substrates (NS). Performance characterization of NS was evaluated by atomic force microscope and X‐ray photoelectron spectroscopy. Urine exfoliated cells were captured by NS and then immunostained to detect urinary tumor cells (UTCs), which was called UTC assay. The receiver operating characteristic (ROC) curve, area under ROC curve (AUC), and Youden index were used to find the cutoff value of UTC assay. ROC analysis and McNemar test were used to compare the diagnostic accuracy of UTC assay with cytology. Kappa test was used to analyze the agreement of UTC assay and cytology with pathological diagnosis.ResultsNanostructured substrates had good cell binding yields of nucleated cells and tumor cells. CK20+CD45−CD11b− cells were considered as UTCs. UTC number ≥ 1 per sample could be considered as a positive result. By AUC and Kappa analysis, UTC assay showed good performance in UC detection. McNemar test demonstrated that UTC assay had a superior sensitivity even in low‐grade subgroup and a similar specificity compared to cytology in UC diagnosis.ConclusionsNanostructured substrates could be used to enrich the exfoliated cells from urine samples. UTC assay with NS has the potential to play a role in UC detection. The value of this assay still needs additional validation by large, multi‐center studies.

Diagnostic performance of an immunoassay based on urine exfoliated cell enrichment nanotechnology for upper tract urothelial carcinoma: a retrospective, monocentric study

“…The supporting software was used to analyze and count cells. The cut-off value of UTC assay is identical to that in our previous report, considering urine tumor cell number ≥ 1 per sample as a positive result [ 9 ].…”

“…However, accurate diagnosis is still challenging owing to the difficult accessibility of the upper tract, causing clinical decision-making intractable, especially when radiologic imaging or biopsies are inconclusive or not diagnostic. We have previously identified a urine-based non-invasive UTC assay which detects bladder UC with high sensitivity and specificity [ 9 ]. Accordingly, this study aimed to assess the value of the UTC assay as a diagnostic tool for UTUC.…”

“…Previously, we developed and validated a non-invasive urine assay that accurately detected UC in bladder, namely “UTC assay”. The UTC assay using voided urine samples showed 80.4% sensitivity and 97.3% specificity for the detection of UC [ 9 ]. Since the bladder UC and UTUC share histologically and cytologically similarities [ 5 ], we hypothesized that UTC assay could be a useful tool for UTUC diagnosis.…”

Evaluation of the value of Preoperative Sialic Acid Levels in Diagnosis and Localization of Urothelial Tumors

“…Therefore, early detection and diagnose have an important significance to the prognosis and quality of life of patients with UC. Cytological experiments and biopsies under an ureteroscope and a bladder scope are currently the two main means of UC early detection and diagnosis [5]. However, use of the ureteroscope and bladder scope not only is invasive but also has a high rate of missed diagnosis, especially for UC in situ and upper tract urothelial carcinoma (UTUC) [5,6].…”

“…Cytological experiments and biopsies under an ureteroscope and a bladder scope are currently the two main means of UC early detection and diagnosis [5]. However, use of the ureteroscope and bladder scope not only is invasive but also has a high rate of missed diagnosis, especially for UC in situ and upper tract urothelial carcinoma (UTUC) [5,6]. Therefore, the search for a specific, highly sensitive marker, that can predict the biological behavior of UC, as an index for predicting tumor progression and metastasis, in order to guide clinical diagnosis and therapy is particularly important.…”