A novel fluorometric aptasensor based on carbon nanocomposite for sensitive detection of Escherichia coli O157:H7 in milk

• Published in: Journal of dairy science Vol. 103; no. 9; pp. 7879 - 7889
• Main Authors: Yang, Tao, Yang, Xinyan, Guo, Xiaojie, Fu, Shiqian, Zheng, Jiapeng, Chen, Sihan, Qin, Xue, Wang, Zhenghui, Zhang, Dongyan, Man, Chaoxin, Jiang, Yujun
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.09.2020
• Subjects: Animals; aptasensor; Biosensing Techniques; Escherichia coli O157 - isolation & purification; Escherichia coli O157:H7; Fluorometry - methods; Fluorometry - veterinary; Food Microbiology; milk; Milk - microbiology; Nanocomposites; Nanotubes, Carbon; Reproducibility of Results; milk; nanocomposites; Escherichia coli O157:H7; aptasensor
• ISSN: 0022-0302; 1525-3198
• DOI: 10.3168/jds.2020-18344

Escherichia coli O157:H7 is an extremely serious foodborne pathogen accounting for a vast number of hospitalizations. In this system, a simple, rapid, and safe compound method was developed based on carbonyl iron powder (CIP) and multiwalled carbon nanotubes (MWCNT). Then, the CIP@MWCNT-based aptasensor was constructed by strong π-stacking between nanocomposite and aptamer, single-strand DNA, causing fluorescent quenching of the dye-labeled aptamer. The restoration of dye fluorescence could be achieved when aptamer came off the surface of the CIP@MWCNT nanocomposite due to the presence of target bacteria. To the best of our knowledge, this fabrication of magnetic carbon nanotubes without irritating and corrosive reagents is described for the first time. The sensing platform was also an improvement on the conventional formation of the aptasensor between carbon materials and DNA aptamer. The nanocomposite was verified by diverse characterization of zeta potential, Fourier-transform infrared spectroscopy, transmission electron microscopy, and energy dispersive x-ray analysis. The CIP@MWCNT-based aptasensor was an effective nanoplatform for quantitative detection of E. coli O157:H7, and was measured to have high specificity, good reproducibility, and strong stability. The aptasensor's capacity to quantify E. coli O157:H7 was as low as 7.15 × 103 cfu/mL in pure culture. The detection limit of E. coli O157:H7 was 3.15 × 102 cfu/mL in contaminated milk after 1 h of pre-incubation. Hence, the developed assay is a new possibility for effective synthesis of nanocomposites and sensitive tests of foodborne pathogens in the dairy industry.