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Bifunctional linker-based immunosensing for rapid and visible detection of bacteria in real matrices.
Biosens Bioelectron. 2018 Feb 15; 100:389-395.BB

Abstract

Detection of pathogens present in food and water is essential to help ensure food safety. Among the popular methods for pathogen detection are those based on culture and colony-counting and polymerase chain reaction (PCR). However, the time-consuming nature and/or the need for sophisticated instrumentation of those methods limit their on-site applications. We have developed a rapid and highly sensitive immunosensing method for visible detection of bacteria in real matrices based on the aggregation of AuNPs without requiring any readout device. We use biotinylated anti-bacteria antibodies as bifunctional linkers (BLs) to mediate the aggregation of streptavidin-functionalized gold nanoparticles (st-AuNPs) to produce visually recognizable color change, due to surface plasmon resonance (SPR), which occurs in about 30min of total assay time when the sample is mildly agitated or within three hours in quiescent conditions. The aggregation of st-AuNPs, which produces the indication signal, is achieved very differently than in visual detection methods reported previously and hence affords ultrahigh sensitivity. While BLs can both bind to the target and crosslink st-AuNPs, their latter function is essentially disabled when they bind to the target bacteria. By varying the amount of st-AuNPs used, we can tailor the assay effectiveness improving limit of detection (LOD) down to 10CFUmL-1 of E. coli and Salmonella. Test results obtained with tap water, lake water and milk samples show that assay performance is unaffected by matrix effects. Further, in a mixture of live and autoclaved E. coli cells our assay could detect only live cells. Therefore, our BL-based immunosensor is suitable for highly sensitive, rapid, and on-site detection of bacteria in real matrices.

Authors+Show Affiliations

Department of Biological Systems Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.Department of Food Science & Technology, Hoseo University, 79-20 Hoseoro, Asan, Chungnam 336-795, South Korea; Center for Natural Sciences, Hoseo University, 79-20 Hoseoro, Asan, Chungnam 336-795, South Korea.Department of Agricultural Biotechnology, Seoul National University, 1 Gwanakro, Gwanakgu, Seoul 151-921, South Korea.Department of Agricultural Biotechnology, Seoul National University, 1 Gwanakro, Gwanakgu, Seoul 151-921, South Korea; Center for Food and Bioconvergence, Seoul National University, 1 Gwanakro, Gwanakgu, Seoul 151-921, South Korea; Research Institute of Agriculture and Life Sciences, Seoul National University, 1 Gwanakro, Gwanakgu, Seoul 151-921, South Korea.Department of Biological Systems Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA. Electronic address: guna@wisc.edu.

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

28954255

Citation

You, Youngsang, et al. "Bifunctional Linker-based Immunosensing for Rapid and Visible Detection of Bacteria in Real Matrices." Biosensors & Bioelectronics, vol. 100, 2018, pp. 389-395.
You Y, Lim S, Hahn J, et al. Bifunctional linker-based immunosensing for rapid and visible detection of bacteria in real matrices. Biosens Bioelectron. 2018;100:389-395.
You, Y., Lim, S., Hahn, J., Choi, Y. J., & Gunasekaran, S. (2018). Bifunctional linker-based immunosensing for rapid and visible detection of bacteria in real matrices. Biosensors & Bioelectronics, 100, 389-395. https://doi.org/10.1016/j.bios.2017.09.033
You Y, et al. Bifunctional Linker-based Immunosensing for Rapid and Visible Detection of Bacteria in Real Matrices. Biosens Bioelectron. 2018 Feb 15;100:389-395. PubMed PMID: 28954255.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Bifunctional linker-based immunosensing for rapid and visible detection of bacteria in real matrices. AU - You,Youngsang, AU - Lim,Seokwon, AU - Hahn,Jungwoo, AU - Choi,Young Jin, AU - Gunasekaran,Sundaram, Y1 - 2017/09/20/ PY - 2017/04/05/received PY - 2017/08/27/revised PY - 2017/09/18/accepted PY - 2017/9/28/pubmed PY - 2018/6/26/medline PY - 2017/9/28/entrez KW - Bifunctional linker KW - Gold nanoparticles KW - Immunoassay KW - Real matrix KW - Streptavidin-biotin binding KW - Visible detection SP - 389 EP - 395 JF - Biosensors & bioelectronics JO - Biosens Bioelectron VL - 100 N2 - Detection of pathogens present in food and water is essential to help ensure food safety. Among the popular methods for pathogen detection are those based on culture and colony-counting and polymerase chain reaction (PCR). However, the time-consuming nature and/or the need for sophisticated instrumentation of those methods limit their on-site applications. We have developed a rapid and highly sensitive immunosensing method for visible detection of bacteria in real matrices based on the aggregation of AuNPs without requiring any readout device. We use biotinylated anti-bacteria antibodies as bifunctional linkers (BLs) to mediate the aggregation of streptavidin-functionalized gold nanoparticles (st-AuNPs) to produce visually recognizable color change, due to surface plasmon resonance (SPR), which occurs in about 30min of total assay time when the sample is mildly agitated or within three hours in quiescent conditions. The aggregation of st-AuNPs, which produces the indication signal, is achieved very differently than in visual detection methods reported previously and hence affords ultrahigh sensitivity. While BLs can both bind to the target and crosslink st-AuNPs, their latter function is essentially disabled when they bind to the target bacteria. By varying the amount of st-AuNPs used, we can tailor the assay effectiveness improving limit of detection (LOD) down to 10CFUmL-1 of E. coli and Salmonella. Test results obtained with tap water, lake water and milk samples show that assay performance is unaffected by matrix effects. Further, in a mixture of live and autoclaved E. coli cells our assay could detect only live cells. Therefore, our BL-based immunosensor is suitable for highly sensitive, rapid, and on-site detection of bacteria in real matrices. SN - 1873-4235 UR - https://www.unboundmedicine.com/medline/citation/28954255/Bifunctional_linker_based_immunosensing_for_rapid_and_visible_detection_of_bacteria_in_real_matrices_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0956-5663(17)30641-3 DB - PRIME DP - Unbound Medicine ER -