Competitive aptasensor for the ultrasensitive multiplexed detection of cancer biomarkers by fluorescent nanoparticle counting

Cancer biomarker quantification in human serum is of great importance for accurate patient diagnosis and informed clinical management. To date, ultrasensitive multiplexed detection of proteins without amplification is still a major challenge. Herein, we proposed a competitive aptasensor strategy for...

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Bibliographic Details
Published in:Analyst (London) 2020-05, Vol.145 (1), p.3612-3619
Main Authors: Pei, Xiaojing, Wu, Xi, Xiong, Jie, Wang, Guohong, Tao, Guangyu, Ma, Yurou, Li, Na
Format: Article
Language:English
Subjects:
Antigens
Aptamers, Nucleotide - genetics
Aptamers, Nucleotide - metabolism
Base Sequence
Biomarkers
Biomarkers, Tumor - analysis
Biomarkers, Tumor - blood
Biomarkers, Tumor - metabolism
Biosensing Techniques - methods
Business competition
Cancer
Deoxyribonucleic acid
Diagnosis
DNA
Fluorescence
Fluorescent Dyes - chemistry
Humans
Limit of Detection
Multiplexing
Nanocomposites
Nanoparticles
Nanoparticles - chemistry
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Summary:Cancer biomarker quantification in human serum is of great importance for accurate patient diagnosis and informed clinical management. To date, ultrasensitive multiplexed detection of proteins without amplification is still a major challenge. Herein, we proposed a competitive aptasensor strategy for ultrasensitive multiplexed cancer biomarker detection by fluorescent nanoparticle (FNP) counting. The sequences are designed such that the binding abilities of linker DNA (L-DNA) with DNA-functionalized FNPs (DNA-FNPs) and aptamer are comparable. As long as one target binds with one molecule of aptamer, a signalling FNP forms a sandwich-structured nanocomposite, which was subsequently observed and enumerated with a fluorescence microscope. This 1 : 1 target-to-signal FNP production assured an improved sensitivity, benefiting from the reasonably good brightness and photostability of FNPs. For both singleplexed and multiplexed detection, this proposed strategy achieved an approximately 1000-fold improved limit of detection than the conventional method with the detection volume of 3.2 μL. Notably, the results for carcinoembryonic antigen (CEA) detection obtained directly from 9 human serum samples (colorectal/lung/healthy individuals) were consistent with that obtained by ELISA, showing potential application in clinical diagnosis. Multiplexed detection by fluorescent nanoparticle counting based on competitive mode.
ISSN:0003-2654
1364-5528
DOI:10.1039/d0an00239a