title: Nicking enzyme-assisted biosensor for Salmonella enteritidis detection based on fluorescence resonance energy transfer.
abstract: Salmonella enteritidis (S. enteritidis) outbreaks continue to occur, and have increased public awareness of this pathogen. Nicking endonuclease Nb.BbvC I is widely used for the detection of biomolecules and displays activity for specific double-stranded DNA (dsDNA). In this study, we developed a biosensor to detect S. enteritidis based on fluorescence resonance energy transfer (FRET) using nicking enzyme and carbon nanoparticles (CNPs). Because of the quenching effect of black hole quencher 1 (BHQ 1), the CNPs do not fluoresce in the reaction system. When the target bacteria are added, the nicking enzyme recognizes and cleaves the dsDNA fabricated by the interaction between probe and target. As a result, the CNPs dissociate from BHQ 1 and emit strong fluorescence. Using the nicking enzyme, the fluorescence signals of the biosensor are greatly amplified. The biosensor exhibited a linear relationship with the concentration of S. enteritidis ranging from 10(2) to 3 × 10(3)CFU/mL in water and from 1.5 × 10(2) to 3 × 10(3)CFU/mL in milk. The present results indicate that our FRET-based detection system can be widely employed for the effective detection of pathogens.
authors.affiliation: Department of Microbiology, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Song
authors.affiliation: Department of Microbiology, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China; College of Life Sciences, Central-South University, Changsha, Hunan 410008, China.
authors.forename: Wenkai
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Microbiology, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China.
authors.forename: Yingfen
authors.initials: Y
authors.lastname: Duan
authors.affiliation: Department of Microbiology, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China.
authors.forename: Zhongjie
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Microbiology, College of Life Sciences, Hunan Normal University, Changsha, Hunan 410081, China. Electronic address: dengle@hunnu.edu.cn.
authors.forename: Le
authors.initials: L
authors.lastname: Deng
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D053904:DNA Breaks, Single-Stranded; D003850:Deoxyribonuclease I / Q000737:chemistry* / Q000493:pharmacokinetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation*; D015203:Reproducibility of Results; D012477:Salmonella enteritidis / Q000302:isolation & purification* / Q000502:physiology*; D012680:Sensitivity and Specificity
chemical_list: D003850:Deoxyribonuclease I
keywords: Biosensor; CNTs; Carbon nanoparticles; Fluorescence signal amplification; Nicking enzyme; Salmonella enteritidis detection