Silver Nanoparticle-Based Ultrasensitive Chemiluminescent Detection of DNA Hybridization and Single-Nucleotide Polymorphisms

• Published in: Analytical chemistry (Washington) 2006-06, Vol.78 (11), p.3738-3744
• Main Authors: Liu, Cheng-Hui, Li, Zheng-Ping, Du, Bao-An, Duan, Xin-Rui, Wang, Yu-Cong
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chemistry; Deoxyribonucleic acid; DNA; DNA - analysis; DNA - chemistry; DNA - genetics; DNA Probes - chemistry; DNA Probes - ultrastructure; Dna, deoxyribonucleoproteins; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Hybridization; Hydrogen-Ion Concentration; Luminescent Measurements - methods; Microscopy, Electron, Transmission; Nanoparticles; Nanoparticles - chemistry; Nanoparticles - ultrastructure; Nucleic acids; Polymorphism; Polymorphism, Single Nucleotide - genetics; Sensitivity and Specificity; Silver; Silver - chemistry; Spectrometric and optical methods; Temperature
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac0522409

A new nanoparticle-based chemiluminescent (CL) method has been developed for the ultrasensitive detection of DNA hybridization. The assay relies on a sandwich-type DNA hybridization in which the DNA targets are first hybridized to the captured oligonucleotide probes immobilized on polystyrene microwells and then the silver nanoparticles modified with alkylthiol-capped oligonucleotides are used as probes to monitor the presence of the specific target DNA. After being anchored on the hybrids, silver nanoparticles are dissolved to Ag+ in HNO3 solution and sensitively determined by a coupling CL reaction system (Ag+−Mn2+−K2S2O8−H3PO4−luminol). The combination of the remarkable sensitivity of the CL method with the large number of Ag+ released from each hybrid allows the detection of specific sequence DNA targets at levels as low as 5 fM. The sensitivity increases 6 orders of magnitude greater than that of the gold nanoparticle-based colorimetric method and is comparable to that of surface-enhanced Raman spectroscopy, which is one of the most sensitive detection approaches available to the nanoparticle-based detection for DNA hybridization. Moreover, the perfectly complementary DNA targets and the single-base mismatched DNA strands can be evidently differentiated through controlling the temperature, which indicates that the proposed CL assay offers great promise for single-nucleotide polymorphism analysis.