Τομέας Κλινικοεργαστηριακός
Library of the School of Health Sciences

2015-03-11

2015

Περικλέους Περικλής

Ευστάθιος Ευσταθόπουλος, Νικόλαος Νικητέας, Μαρία Γαζούλη

Ανάπτυξη νανοανιχνευτών για τη στοχευμένη μοριακή ανίχνευση και απεικόνιση των μικρομεταστάσεων

Greek

Development of a quantum dot labelled magnetic immunoassay method for the detection of circulating tumour cells and micrometastases

Aim-Background
To evaluate the incorporation of cadmium selenide quantum dots (QDs) in the
detection of colorectal cancer (CRC) circulating tumour cell (CTC) surface
antigens.
Material and Methods
The principle of this assay is based upon the separation of CTCs from body
fluids using magnetic beads (MBs) coupled with specific antibody biomarkers,
such as the Epithelial cell Adhesion Molecule Antibody (EpCAM) and Cytokeratin
19 (CK19) antibody. The detection signal was acquired from the fluorescence
signal of QDs. To evaluate 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 samples. Peripheral blood samples were obtained from
9 CRC patients and 5 healthy donors who gave their informed consent to their
inclusion in the study. Peripheral venous blood was sampled immediately after
CRC patients had received general anaesthesia, moments before surgery. In all
patients, an intravenous cannula was used to collect blood into 7 ml
vacutainers containing sodium ethylenediaminetetraacetic acid (EDTA). The first
7 ml of blood was discarded in order to reduce the risk of contamination from
skin epithelial cells. Subsequently, 30 ml samples were collected at one-minute
intervals (five 6 ml aliquots per 30 ml sample).
Results
The limit of detection (LOD) of the methodology described in the present study
was determined at 10 DLD-1 cells/ml of sample as fluorescence measured with a
spectrofluorometer. Fluorescence activated cell sorting (FACS) analysis and
Real Time RT-PCR were also used to evaluate the performance of the method.
Ultimately, in comparison to other methods currently in use, we developed a
simple, sensitive, and more cost-effective method for the detection of CRC CTCs
in human samples. Results were accomplished using magnetic bead isolation and
subsequent QD fluorescence detection.
Conclusion
The present method can be readily adjusted to target a variety of proteins of
either the CTCs or the host.

Quantum dots, Micrometastases, Circulating tumour cells, Cancer, Nanoprobes

No

0

Yes

223

106, x

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https://pergamos.lib.uoa.gr/uoa/dl/object/1305657