Journal
BIOSENSORS & BIOELECTRONICS
Volume 85, Issue -, Pages 734-739Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2016.05.057
Keywords
FITC; Signal amplification; Dual lateral flow immunoassay; Escherichia coli O157:H7
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Funding
- National Natural Science Foundation of China [31371776]
- Capability Construction Program of Science and Technology Commission of Shanghai [13430502400]
- Science and Technology Innovation Plan of Shanghai: Yangtze River Delta Joint Research [15395810900]
- China Postdoctoral Science Foundation [2015M581637]
- Cultivation Fund for National Project of University of Shanghai for Science and Technology [16HJPY-QN09]
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A pattern of signal amplification lateral flow immunoassay (LFIA) for pathogen detection, which used fluorescein isothiocyanate (FITC) labeled antigen and antibody for dual FITC-LFIA was developed. Escherichia coli O157:H7 (E.coli O157:H7) was selected as the model analyte. In the signal amplification LFIA method, FITC was mixed with sample culture medium, with the presence of E.coli O157:H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation, creating a fluorescent antigen probe. This antigen probe was added to LFIA, which already contained E.coli O157:H7 monoclonal antibodies-FITC (McAb-E.coli O157:H7-FITC) dispersed in the conjugate pad. Another E.coli O157:H7 McAb was the test line, and goat anti-mouse IgG antibody was the control line in nitrocellulose (NC) membrane. The visual limit of detection (LOD) of the strip for qualitative detection was 10(5) CFU/mL while the LOD for semi-quantitative detection could down to 104 CFU/mL by using scanning reader. Signal amplification LFIA was perfectly applied to the detection of food samples with E.coli O157:H7. The LOD was substantially improved to 1 CFU/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 8 and 8 h respectively. The results of this method was more sensitive by 10-fold than the conventional colloidal gold (CG) based strips and comparable to the traditional ELISA. This simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring food-borne pathogens in food-safety testing. (C) 2016 Elsevier B.V. All rights reserved.
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