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KMID : 1007520210300081129
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Food Science and Biotechnology
2021 Volume.30 No. 8 p.1129 ~ p.1138
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A detection method of Escherichia coli O157:H7 based on immunomagnetic separation and aptamers-gold nanoparticle probe quenching Rhodamine B’s fluorescence
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Lian Fengnan
Wang Dan
Yao Shuo
Ge Lirui
Wang Yue
Zhao Yuyi
Zhao Jinbin
Song Xiuling
Zhao Chao
Li Jinhua
Liu Yajuan
Jin Minghua
Xu Kun
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Abstract
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This research aimed to detect Escherichia coli O157:H7 in milk based on immunomagnetic probe separation technology and quenching effect of gold nanoparticles to Rhodamine B. Streptavidin-modified magnetic beads (MBs) were combined with biotin-modified antibodies to capture E. coli O157:H7 specifically. Gold nanoparticle (AuNPs) was incubated with sulfhydryl-modified aptamers (SH-Aptamers) to obtain the Aptamers-AuNPs probe. After magnetic beads captured target bacteria and formed a sandwich structure with the gold nanoprobe, Rhodamine B was added into complex to obtain fluorescent signal changes. Our results demonstrated that the established method could detect E. coli O157:H7 in the range of 101?107 CFU/mL, and the limit of detection (LOD) was 0.35 CFU/mL in TBST buffer (pH?=?7.4). In milk simulation samples, the LOD of this method was 1.03 CFU/mL. Our research provides a promising approach on the detection of E. coli O157:H7.
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KEYWORD
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Immunomagnetic, Gold nanoparticle, Aptamer, Rhodamine B, Foodborne pathogen
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