Quantum-dot-based suspension microarray for multiplex detection of lung cancer markers: preclinical validation and comparison with the Luminex xMAP system.
Fiche publication
Date publication
mars 2017
Journal
Scientific reports
Auteurs
Membres identifiés du Cancéropôle Est :
Pr NABIEV Igor
Tous les auteurs :
Bilan R, Ametzazurra A, Brazhnik K, Escorza S, Fernández D, Uríbarri M, Nabiev I, Sukhanova A
Lien Pubmed
Résumé
A novel suspension multiplex immunoassay for the simultaneous specific detection of lung cancer markers in bronchoalveolar lavage fluid (BALF) clinical samples based on fluorescent microspheres having different size and spectrally encoded with quantum dots (QDEM) was developed. The designed suspension immunoassay was validated for the quantitative detection of three lung cancer markers in BALF samples from 42 lung cancer patients and 10 control subjects. Tumor markers were detected through simultaneous formation of specific immune complexes consisting of a capture molecule, the target antigen, and biotinylated recognition molecule on the surface of the different QDEM in a mixture. The immune complexes were visualized by fluorescently labeled streptavidin and simultaneously analyzed using a flow cytometer. Preclinical validation of the immunoassay was performed and results were compared with those obtained using an alternative 3-plex immunoassay based on Luminex xMAP technology, developed on classical organic fluorophores. The comparison showed that the QDEM and xMAP assays yielded almost identical results, with clear discrimination between control and clinical samples. Thus, developed QDEM technology can become a good alternative to xMAP assays permitting analysis of multiple protein biomarkers using conventional flow cytometers.
Mots clés
Adult, Aged, Aged, 80 and over, Biomarkers, Tumor, metabolism, Bronchoalveolar Lavage Fluid, Female, Flow Cytometry, Humans, Immunoassay, methods, Lung Neoplasms, diagnosis, Male, Microspheres, Middle Aged, Multivariate Analysis, Quantum Dots, chemistry, Reproducibility of Results
Référence
Sci Rep. 2017 03 16;7:44668