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Objective: To develop and validate a two-step enrichment-identification system for circulating tumor cells (CTCs) by improving the isolation by size of tumor cells (ISET) technology and combining it with an immunocytochemistry (ICC) technique. Methods: The peripheral blood (PB) samples from 23 patients with late stage (III/IV) tumors was used to validate the ISET technology used in combination with the identification techniques of ICC and wax block-immunohistochemistry (IHC). Comparison of the diagnostic performance of the ISET-ICC system with that of CTC-biopsy and CellSearchTM methods Methods: The peripheral blood (PB) samples from 23 patients with late stage (III/IV) tumors was used to validate the ISET technology used in combination with the identification techniques of ICC and wax block-immunohistochemistry (IHC). Comparison of the diagnostic performance of the ISET-ICC system with that of CTC-biopsy and CellSearchTM methods was performed using samples from 74 patients with six different types of cancer. Results: The two-step enrichment-identification ISET filtration-based method combined with ICC detection output a CTC-positive rate of 65.2%. We also showed that the ISET-ICC system can be combined with the cell wax block-IHC method to increase the reliability of EMT-CTC identification. The developed ISET-ICC technology showed a significantly higher CTC-positive rate (17.6%; 13/74) for the study cohort of 74 cases with various cancer types than the only Federal Drug Administration (FDA)-approved CellSearchTM method (12.2%; 9/74; 2 = 10.21, p = 0.007). In addition, the accuracy of the different CTC diagnostic methods is tumor-type dependent. Moreover, we noted a correlation between the CTC-positive rate and the tumor stage. Conclusion: We have established an improved ISET-ICC system that can successfully detect different CTC types in various cancer types that shows a higher accuracy compared with the CellSearchTM system.

journal of cancer science and clinical therapeutics

Detection of Circulating Tumor Cells in Peripheral Blood by A Novel Two-Step Tumor Cell Enrichment-Identification System that Combines the Isolation by Size of Tumor Cells Technology and the Immunofluorescence Technique