Canine lower urinary tract neoplasia is a clinically important disease process with high mortality due to it typically having a late stage diagnosis and a poorly durable response to treatment. Non-invasive diagnostic techniques (e.g. dipstick test, urine cytology) have poor diagnostic value, while more invasive tests (e.g. cystoscopy and biopsy) are costly, often require general anaesthesia, and carry the potential to spread the disease to other sites. Here we have developed a quantitative cytological analysis method (SERS cytology) for the identification of cancerous transitional cells in urine, based on the use of multiplexed surface-enhanced Raman spectroscopy (SERS) and chemometrics analysis. The assay uses silver nanoparticle-based SERS biotags (SBTs) carrying the peptide PLZ4 (amino acid sequence cQDGRMGFc) that targets malignant transitional cells. Cells are imaged by bright field microscopy, binned according to their morphological characteristics, and those showing the hallmarks of transitional cells are analyzed by SERS-cytology, obtaining single-cell data. Transitional neoplastic cells are reliably identified by looking at the ratio of two tags -- the SERS signal produced by a bladder cancer marker (PLZ4) versus that of an internal control (a SERS marker that binds to both healthy and TCC cells equally well). Samples derived from patients diagnosed with TCC showed significantly higher mean PLZ4/TAT ratios (&gt; 1.2) than their healthy counterparts (p&lt;0.05). The assay specificity was determined to be 83%, and sensitivity 100%, improving over the sensitivity of standard cytology. The cellular fraction of urine sediment is a highly complex medium, characterized by the presence of several types of cells. Here we show that multiplexed SERS, coupled with chemometric analysis can successfully identify transitional carcinoma cells (TCC) quantitatively in clinical urine samples.

Screening for canine transitional cell carcinoma (TCC) by SERS-based quantitative urine cytology

256th National Meeting (2018)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.