Portable dual-aptamer microfluidic chip biosensor for Bacillus cereus based on aptamer tailoring and dumbbell-shaped probes

Ziqi Zhou; Xinyue Lan; Longjiao Zhu; Yangzi Zhang; Kehan Chen; Wenqiang Zhang; Wentao Xu

doi:10.1016/j.jhazmat.2022.130545

Portable dual-aptamer microfluidic chip biosensor for Bacillus cereus based on aptamer tailoring and dumbbell-shaped probes

J Hazard Mater. 2023 Mar 5:445:130545. doi: 10.1016/j.jhazmat.2022.130545. Epub 2022 Dec 5.

Authors

Ziqi Zhou¹, Xinyue Lan¹, Longjiao Zhu², Yangzi Zhang¹, Kehan Chen³, Wenqiang Zhang⁴, Wentao Xu⁵

Affiliations

¹ Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, Beijing 100193, China; Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety) (MOA), College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China.
² Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, Beijing 100193, China.
³ Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, Beijing 100193, China; Department of Mechanical Design and Manufacturing, College of Engineering, China Agricultural University, Beijing 100083, China.
⁴ Department of Mechanical Design and Manufacturing, College of Engineering, China Agricultural University, Beijing 100083, China.
⁵ Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, Beijing 100193, China. Electronic address: xuwentao@cau.edu.cn.

PMID: 36493638
DOI: 10.1016/j.jhazmat.2022.130545

Abstract

As food-borne pathogens, Bacillus cereus not only produce toxins that contaminate food and threaten human health, but also rely on spores to resist extreme environments. At present, the detection of B. cereus is still at the genome level and it is not easily distinguished from other Bacilli of the same group. Herein, we obtained the aptamers of B. cereus in different phases through Cell-SELEX technology. Then, through step-by-step tailoring and molecular docking, the two best performing aptamers were ascertained and the interaction revealed between the repeated G bases in the aptamer and the polar amino acids in the α-helix of the epiprotein. Based on these aptamers, a multifunctional dumbbell-shaped probe and an ultrasensitive microfluidic chip biosensor were designed. Tests showed that the novel sensor is able to complete detection within 1 h with a limit of detection (LOD) of 9.27 CFU/mL. Moreover, the sensor can be used in complex food environments, such as milk and rice, is able to detect both vegetative cells and spores, and it can also distinguish B. thuringiensis from the same flora. This study can provide a reference for the future development of food-borne pathogenic bacteria aptamer selecting, target interaction analysis, detection methods and equipment.

Keywords: Aptamer; Bacillus cereus; Microfluidic biosensor; SELEX; Tailoring.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacillus cereus
Bacillus*
Biosensing Techniques*
Humans
Microfluidics
Molecular Docking Simulation
Oligonucleotides

Substances

Oligonucleotides