Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 411, Issue 19, Pages 4569-4576Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-018-1422-y
Keywords
Biosensor; Cascade; Label-free; Hairpin DNA; Exonuclease III
Funding
- National Natural Science Foundation of China [51572044, 21475018]
- Fundamental Research Funds for the Central Universities [N160504010, N170507001]
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A simple fluorescence biosensor is developed based on the enzyme-assisted cascade amplification strategy. The amplification system consists of a hairpin-structure DNA (H-DNA) and exonuclease III. The target DNA can hybridize with the H-DNA and initiate exonuclease III-assisted target recycling amplification to generate abundant G-rich DNA (G-DNA). One region of G-DNA is designed to possess the same sequence as target DNA. Thus, the G-DNA can also hybridize with H-DNA and initiate the digestion of H-DNA. The cascade strategy in this amplification system causes the concentration of G-DNA to grow exponentially. The fluorescence intensity of N-methylmesoporphyrin IX (NMM) is highly enhanced due to the formation of G-quadruplex configuration. Under optimal conditions, the cascade system could achieve an admirable sensitivity with a detection limit of 52fM for HIV DNA, and guarantees a satisfactory specificity. Moreover, the cascade system could be implemented for other target DNA detections by substituting the recognition region of the H-DNA. In this way, a detection limit of 65fM for HBV DNA could be achieved by the cascade system. The target DNA analysis in a real serum sample further indicates that this biosensor has potential for future application in clinical diagnosis.
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