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A nanoporous membrane-based impedimetric immunosensor for label-free detection of pathogenic bacteria in whole milk.
Biosens Bioelectron. 2013 Jun 15; 44:210-5.BB

Abstract

We introduce a nanoporous membrane based impedimetric immunosensor for the label-free detection of bacterial pathogens in whole milk. A simple and rapid method to modify a commercially available alumina nanoporous membrane with hyaluronic acid (HA) effectively reduced the non-specific binding of biomolecules and other cells, and permitted successful immobilization of antibodies. Escherichia coli O157:H7, one of the most harmful food-borne pathogenic bacteria, was tested as a model pathogen in this study. The ionic impedance of electrolytes through nanopores, due to antibody-pathogen interactions, was monitored by impedance spectra and analyzed by normalized impedance change (NIC). The regression equation for the NIC at 1 kHz versus concentration of E. coli O157:H7 (10-10(5)cfu/ml) was obtained, and the detection limit found to be as low as 10 cfu/ml. In addition, the proposed immunosensor was successfully used for the detection of E. coli O157:H7 in whole milk samples with the detection limit as low as 83.7 cfu/ml with 95% probability. The specificity of the immunosensor was also demonstrated using non-target bacteria, including Staphylococcus aureus, Bacillus cereus, and non pathogenic E. coli DH5α. This study shows that a HA-functionalized nanoporous membrane-based impedimetric sensor is capable of detecting pathogenic bacteria in whole milk without any pretreatment. This is a significant step for evaluating the safety of food and environmental samples and other medical diagnostics.

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Authors+Show Affiliations

Joung CK
Institute of Life Sciences and Resources & Department of Food Science and Biotechnology, College of Life Sciences, Kyung Hee University, Yongin, Republic of Korea.
Kim HN
No affiliation info available
Lim MC
No affiliation info available
Jeon TJ
No affiliation info available
Kim HY
No affiliation info available
Kim YR
No affiliation info available

MeSH

Aluminum OxideAnimalsAntibodies, ImmobilizedBacillus cereusElectric ImpedanceEquipment DesignEscherichia coli O157Food MicrobiologyHyaluronic AcidImmunoassayLimit of DetectionMembranes, ArtificialMilkPorosityStaphylococcus aureus

Pub Type(s)

Evaluation Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23428735

Citation

Joung, Cho-Kyung, et al. "A Nanoporous Membrane-based Impedimetric Immunosensor for Label-free Detection of Pathogenic Bacteria in Whole Milk." Biosensors & Bioelectronics, vol. 44, 2013, pp. 210-5.
Joung CK, Kim HN, Lim MC, et al. A nanoporous membrane-based impedimetric immunosensor for label-free detection of pathogenic bacteria in whole milk. Biosens Bioelectron. 2013;44:210-5.
Joung, C. K., Kim, H. N., Lim, M. C., Jeon, T. J., Kim, H. Y., & Kim, Y. R. (2013). A nanoporous membrane-based impedimetric immunosensor for label-free detection of pathogenic bacteria in whole milk. Biosensors & Bioelectronics, 44, 210-5. https://doi.org/10.1016/j.bios.2013.01.024
Joung CK, et al. A Nanoporous Membrane-based Impedimetric Immunosensor for Label-free Detection of Pathogenic Bacteria in Whole Milk. Biosens Bioelectron. 2013 Jun 15;44:210-5. PubMed PMID: 23428735.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A nanoporous membrane-based impedimetric immunosensor for label-free detection of pathogenic bacteria in whole milk. AU - Joung,Cho-Kyung, AU - Kim,Han-Nah, AU - Lim,Min-Cheol, AU - Jeon,Tae-Joon, AU - Kim,Hae-Yeong, AU - Kim,Young-Rok, Y1 - 2013/01/23/ PY - 2012/10/18/received PY - 2012/12/26/revised PY - 2013/01/09/accepted PY - 2013/2/23/entrez PY - 2013/2/23/pubmed PY - 2013/8/30/medline SP - 210 EP - 5 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 44 N2 - We introduce a nanoporous membrane based impedimetric immunosensor for the label-free detection of bacterial pathogens in whole milk. A simple and rapid method to modify a commercially available alumina nanoporous membrane with hyaluronic acid (HA) effectively reduced the non-specific binding of biomolecules and other cells, and permitted successful immobilization of antibodies. Escherichia coli O157:H7, one of the most harmful food-borne pathogenic bacteria, was tested as a model pathogen in this study. The ionic impedance of electrolytes through nanopores, due to antibody-pathogen interactions, was monitored by impedance spectra and analyzed by normalized impedance change (NIC). The regression equation for the NIC at 1 kHz versus concentration of E. coli O157:H7 (10-10(5)cfu/ml) was obtained, and the detection limit found to be as low as 10 cfu/ml. In addition, the proposed immunosensor was successfully used for the detection of E. coli O157:H7 in whole milk samples with the detection limit as low as 83.7 cfu/ml with 95% probability. The specificity of the immunosensor was also demonstrated using non-target bacteria, including Staphylococcus aureus, Bacillus cereus, and non pathogenic E. coli DH5α. This study shows that a HA-functionalized nanoporous membrane-based impedimetric sensor is capable of detecting pathogenic bacteria in whole milk without any pretreatment. This is a significant step for evaluating the safety of food and environmental samples and other medical diagnostics. SN - 1873-4235 UR - https://www.unboundmedicine.com/medline/citation/23428735/A_nanoporous_membrane_based_impedimetric_immunosensor_for_label_free_detection_of_pathogenic_bacteria_in_whole_milk_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0956-5663(13)00027-4 DB - PRIME DP - Unbound Medicine ER -