Study on Evanescent Wave Fluorescence Aptasensor for Direct and Rapid Detection of Escherichia coli O157∶H7

doi:10.13560/j.cnki.biotech.bull.1985.2019-1132

Abstract

Abstract: By combining the advantages of evanescent wave fluorescence optical fiber biosensor and specific nucleic acid aptamer,a fast and direct method to detect Escherichia coli O157∶H7（E. coli O157∶H7）based on evanescent wave fluorescence principle and its size effect with pathogen was proposed. The basic principle is that when a certain concentration of fluorescence labeled E. coli O157∶H7 aptamer is added to the sample detection cell,the evanescent wave triggers the fluorescence molecules to emit fluorescence,and the evanescent wave all fiber biosensor is applied for the quantitative detection of fluorescence signal. After the fluorescence labeled aptamer is mixed with E. coli O157∶H7,the mixture is added to the sample detection cell. Because the penetration depth of evanescent wave is only 100 nm,the fluorescence molecules labeled aptamer specifically bind with cannot be triggered,which results in the decrease of detected fluorescence signal. The quantitative detection of E. coli O157∶H7 can be achieved by using the proportional relationship between the fluorescence intensity and its concentrations. The results showed that the detection limit of E. coli O157∶H7 was 610 CFU/ml,and the linear detection range was from 1.1×10³ to 1.4 × 10⁷ CFU/mL. The recovery of E. coli O157∶H7 spiked in real samples was 40%-180% and the relative standard deviation was within 10%. The matrix of water sample had no significant effect on the detection of E. coli O157∶H7. Based on the principle of evanescent wave fluorescence and its size effect on pathogens,this universal biosensing method can be applied for the direct and rapid detection of other pathogenic bacteria only using different fluorescence labeled bio-recognition molecules.

Key words: Escherichia coli O157:H7, nucleic acid aptamer, evanescent wave fluorescence, biosensor

FANG Shun-yan, SONG Dan, LIU Yan-ping, XU Wen-juan, LIU Jia-yao, HAN Xiang-zhi, LONG Feng. Study on Evanescent Wave Fluorescence Aptasensor for Direct and Rapid Detection of Escherichia coli O157∶H7[J]. Biotechnology Bulletin, 2020, 36(7): 228-234.

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