Citation: DENG Jian-Kang, NIU Xiao-Juan, LIU Ya-Qing, WANG Shuo. A Ratiometric Fluorescent Biosensor Based on Exonuclease Ⅲ Amplification Strategy for Pathogenic Bacterial Gene Detection[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(5): 743-749. doi: 10.11895/j.issn.0253-3820.181091 shu

A Ratiometric Fluorescent Biosensor Based on Exonuclease Ⅲ Amplification Strategy for Pathogenic Bacterial Gene Detection

¹ State Key Laboratory of Food Nutrition and Safety, Key laboratory of Food Nutrition and Safety (Ministry of Education), College of Food Engineering and Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China;
² School of Medicine, Nankai University, Tianjin 300457, China

Received Date: 3 February 2018
Revised Date: 13 March 2018

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21775108, 21575138).

Food-borne pathogenic bacteria seriously threaten public health. Based on the mechanism of fluorescence resonance energy transfer (FRET), a ratiometric fluorescence biosensor was constructed by integration of Exo III-based signal amplification strategy. The Cy3 labeled R1-DNA firstly hybridized with Cy5 labeled R2-DNA to form duplex of R1/R2. Cy3 showed a low fluorescence response while Cy5 showed a high fluorescence response. The addition of target pathogenic bacterial gene (Lac Z gene) could de-hybridize the R1/R2, resulting in the fluorescence decreasing of Cy5 and the fluorescence recovering of Cy3. Under the assistance of Exo III, the signal change was further amplified. The detection of limit reached as low as 5.29 pmol/L. The linear detection range was from 10 pmol/L to 2000 pmol/L. The developed ratiomtric detection strategy significantly reduced the possibility of false-positive and false-negative detection results.
- Escherichia coli,
- Pathogenic bacterial gene,
- Fluorescence resonance energy transfer,
- Exonuclease Ⅲ,
- Ratiometric fluorescence biosensor
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