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Development of a quantum-dot-labelled magnetic immunoassay method for circulating colorectal cancer cell detection
Authors:Gazouli Maria  Lyberopoulou Anna  Pericleous Pericles  Rizos Spyros  Aravantinos Gerassimos  Nikiteas Nikolaos  Anagnou Nicholas P  Efstathopoulos Efstathios P
Affiliation:Maria Gazouli, Anna Lyberopoulou, Nicholas P Anagnou, Department of Basic Medical Sciences, Laboratory of Biology, School of Medicine, University of Athens, 11527 Athens, Greece.
Abstract:
AIM: To detect of colorectal cancer (CRC) circulating tumour cells (CTCs) surface antigens, we present an assay incorporating cadmium selenide quantum dots (QDs) in these paper.METHODS: The principle of the assay is the immunomagnetic separation of CTCs from body fluids in conjunction with QDs, using specific antibody biomarkers: epithelial cell adhesion molecule antibody, and monoclonal cytokeratin 19 antibody. The detection signal was acquired from the fluorescence signal of QDs. For the evaluation of the performance, the method under study was used to isolate the human colon adenocarcinoma cell line (DLD-1) and CTCs from CRC patients’ peripheral blood.RESULTS: The minimum detection limit of the assay was defined to 10 DLD-1 CRC cells/mL as fluorescence was measured with a spectrofluorometer. Fluorescence-activated cell sorting analysis and Real Time RT-PCR, they both have also been used to evaluate the performance of the described method. In conclusion, we developed a simple, sensitive, efficient and of lower cost (than the existing ones) method for the detection of CRC CTCs in human samples. We have accomplished these results by using magnetic bead isolation and subsequent QD fluorescence detection.CONCLUSION: The method described here can be easily adjusted for any other protein target of either the CTC or the host.
Keywords:Circulating tumor cells   Cancer   Quantum dots   Nanoprobes   Micrometastasis
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