Highly sensitive and selective chemiluminescent imaging for DNA detection by ligation-mediated rolling circle amplified synthesis of DNAzyme

• Published in: Biosensors & bioelectronics Vol. 41; pp. 348 - 353
• Main Authors: Dong, Haifeng, Wang, Chen, Xiong, Yi, Lu, Huiting, Ju, Huangxian, Zhang, Xueji
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 15.03.2013; Elsevier
• Subjects: Biological and medical sciences; Biosensing Techniques - instrumentation; Biosensors; Biotechnology; Chemiluminescent imaging; DNA - analysis; DNA - chemistry; DNA - genetics; DNA, Catalytic - chemistry; DNA, Catalytic - genetics; DNAzyme; Equipment Design; Equipment Failure Analysis; Fundamental and applied biological sciences. Psychology; Ligase; Luminescent Measurements - instrumentation; Methods. Procedures. Technologies; Nucleic Acid Amplification Techniques - methods; Oligonucleotide Array Sequence Analysis - instrumentation; Reproducibility of Results; Rolling circle amplification; Sensitivity and Specificity; Various methods and equipments; DNAzyme; Ligase; Rolling circle amplification; Biosensors; Chemiluminescent imaging; Enzyme; Selectivity; Imagery; Amplification; Ligases; Biosensor; DNA; Detection
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2012.08.050

A highly sensitive DNA biosensing method down to sub-femtomolar level with excellent selectivity was proposed by designing an amplified synthesis of horseradish peroxidase mimicking DNAzyme and introducing the amplified DNAzyme to chemiluminescent (CL) imaging. The amplified synthesis was achieved by combining a target DNA related ligase reaction with rolling circle amplification (RCA), which produced thousands of repeated sequences to bind hemin and form a mass of horseradish peroxidase-mimicing DNAzyme units. The amplification strategy greatly enhanced the CL emission of the luminol–H2O2 system. The genotyping method displayed highly specific biochemistry in allele discrimination. The novel CL imaging strategy based on ligation-mediated RCA synthesis of DNAzyme showed high fidelity in discriminating single-base mismatch and efficiently facilitated signal amplification for sensitive target DNA detection. It could detect DNA ranging from 1×10−15M to 1×10−11M with a detection limit of 0.26fM. The proposed approach provided a robust, cost-efficient, highly sensitive and specific platform for genetic target analysis in bioanalysis and clinic biomedical application. ► This work constructs a highly sensitive chemiluminescent imaging method. ► A novel signal amplification strategy to enhance the sensitivity is proposed. ► Ligation-mediated rolling circle amplification of DNAzyme is introduced to CL imaging. ► This biosensor can detect target DNA down to sub-femtomolar level.