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Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor.
Talanta. 2016 Aug 15; 156-157:42-47.T

Abstract

A simple, one-step, rapid method to detect Escherichia coli O157: H7 (E. coli O157: H7) using a label-free immunofluorescence strip sensor is presented. Fluorescein isothiocyanate (FITC) was added to the sample culture medium to prepare the fluorescent probe for the label-free strip sensor. With the presence of E. coli O157: H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation and maintain good affinity to the monoclonal antibodies (McAb) against E. coli O157: H7. The direct-type immunofluorescence strip sensor was based on the binding between fluorescent bacteria and the unlabeled McAb immobilized at the test line in nitrocellulose membrane (NC membrane) reaction zone. The visual limit of detection (LOD) of the strip for qualitative detection was 10(6)cells/mL while the LOD for semi-quantitative detection could go down to 10(5)cells/mL by using scanning reader. The LOD was substantially improved to 1cells/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 10 and 8h respectively, which was competitive to some current rapid E. coli O157: H7 detection methods. Besides the obvious advantages, including reduced detection time and operation procedures, the results of this method meet the various detection requirements for E. coli O157: H7 and are comparable to the traditional enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich gold-labeled strips. This is the first report of semi-quantitative immunofluorescence strip for directly detecting foodborne pathogen using only one unlabeled antibody. All detections could be achieved in less than 5min. In addition, this simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring foodborne pathogens in food-safety testing.

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Publisher Full Text (DOI)

Authors+Show Affiliations

Song C
School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.
Li J
School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.
Liu J
School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.
Liu Q
School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China. Electronic address: liuq@usst.edu.cn.

MeSH

AnimalsAntibodies, ImmobilizedEquipment DesignEscherichia coli O157Fluorescein-5-isothiocyanateFluorescent Antibody TechniqueFluorescent DyesFood AnalysisFood ContaminationFood MicrobiologyLimit of DetectionMilkReagent Strips

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

27260433

Citation

Song, Chunmei, et al. "Simple Sensitive Rapid Detection of Escherichia Coli O157:H7 in Food Samples By Label-free Immunofluorescence Strip Sensor." Talanta, vol. 156-157, 2016, pp. 42-47.
Song C, Li J, Liu J, et al. Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor. Talanta. 2016;156-157:42-47.
Song, C., Li, J., Liu, J., & Liu, Q. (2016). Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor. Talanta, 156-157, 42-47. https://doi.org/10.1016/j.talanta.2016.04.054
Song C, et al. Simple Sensitive Rapid Detection of Escherichia Coli O157:H7 in Food Samples By Label-free Immunofluorescence Strip Sensor. Talanta. 2016 Aug 15;156-157:42-47. PubMed PMID: 27260433.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simple sensitive rapid detection of Escherichia coli O157:H7 in food samples by label-free immunofluorescence strip sensor. AU - Song,Chunmei, AU - Li,Jianwu, AU - Liu,Jinxin, AU - Liu,Qing, Y1 - 2016/04/29/ PY - 2016/02/28/received PY - 2016/04/09/revised PY - 2016/04/24/accepted PY - 2016/6/5/entrez PY - 2016/6/5/pubmed PY - 2018/2/1/medline KW - Escherichia coli O157:H7 KW - FITC KW - Immunofluorescence KW - Label-free KW - Strip SP - 42 EP - 47 JF - Talanta JO - Talanta VL - 156-157 N2 - A simple, one-step, rapid method to detect Escherichia coli O157: H7 (E. coli O157: H7) using a label-free immunofluorescence strip sensor is presented. Fluorescein isothiocyanate (FITC) was added to the sample culture medium to prepare the fluorescent probe for the label-free strip sensor. With the presence of E. coli O157: H7 in the samples, the bacteria could emit a yellow-green fluorescence after incubation and maintain good affinity to the monoclonal antibodies (McAb) against E. coli O157: H7. The direct-type immunofluorescence strip sensor was based on the binding between fluorescent bacteria and the unlabeled McAb immobilized at the test line in nitrocellulose membrane (NC membrane) reaction zone. The visual limit of detection (LOD) of the strip for qualitative detection was 10(6)cells/mL while the LOD for semi-quantitative detection could go down to 10(5)cells/mL by using scanning reader. The LOD was substantially improved to 1cells/mL of the original bacterial content after pre-incubation of the bread, milk and jelly samples in broth for 10, 10 and 8h respectively, which was competitive to some current rapid E. coli O157: H7 detection methods. Besides the obvious advantages, including reduced detection time and operation procedures, the results of this method meet the various detection requirements for E. coli O157: H7 and are comparable to the traditional enzyme-linked immunosorbent assay (ELISA) and double antibody sandwich gold-labeled strips. This is the first report of semi-quantitative immunofluorescence strip for directly detecting foodborne pathogen using only one unlabeled antibody. All detections could be achieved in less than 5min. In addition, this simple, low-cost and easy to be popularized method served as a significant step towards the development of monitoring foodborne pathogens in food-safety testing. SN - 1873-3573 UR - https://www.unboundmedicine.com/medline/citation/27260433/Simple_sensitive_rapid_detection_of_Escherichia_coli_O157:H7_in_food_samples_by_label_free_immunofluorescence_strip_sensor_ DB - PRIME DP - Unbound Medicine ER -