Detection of Salmonella in Milk Using Enzymatic Precipitation
Enhanced SPR Immunosensor

a 2017

Detection of Salmonella in Milk Using Enzymatic Precipitation Enhanced SPR Immunosensor

PASTUCHA, Matěj, Zdeněk FARKA, Tomáš JUŘÍK and Petr SKLÁDAL

Basic information

Original name

Detection of Salmonella in Milk Using Enzymatic Precipitation Enhanced SPR Immunosensor

Authors

PASTUCHA, Matěj, Zdeněk FARKA, Tomáš JUŘÍK and Petr SKLÁDAL

Edition

CEITEC PhD Retreat II, 2017

Other information

Language

English

Type of outcome

Konferenční abstrakt

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Organization unit

Central European Institute of Technology

ISBN

978-80-210-8550-3

Keywords in English

Surface Plasmon Resonance; Immunosensor; Salmonella; Enzymatic Precipitation; Food Safety

Abstract

V originále

Foodborne bacterial pathogens represent a serious threat for public health. Recent outbreaks have demonstrated the need for rapid and reliable screening techniques. As an alternative to the conventional methods, various biosensors have been developed. They offer fast, sensitive, specific, robust and cost effective analysis and have a potential for point-of-care application. In this work, we have developed a surface plasmon resonance (SPR) immunosensor for the detection of Salmonella Typhimurium in powdered milk. In addition to a conventional label free SPR assay, an enzymatic enhancement step was incorporated. The method did not require any pre enrichment, making the assay fast and simple. After Salmonella was bound by capture antibody immobilized on the sensor surface, solution of detection antibody labelled by horseradish peroxidase was injected. The bound enzymatic label in turn catalyzed the precipitation of 4-chloro-1-naphthol to insoluble benzo-4-chlorocyclohexadienone. The formation of precipitate on the sensor surface provided substantial signal enhancement. The precipitation-based assay format improved the sensitivity 40 times compared to the label-free format. The limit of detection of 10^2 CFU·mL-1 in buffer and 10^3 CFU·mL–1 in milk was achieved and the whole analysis time was below 60 min. The interaction of the bacteria with the sensor surface and the formation of the precipitate were studied in detail by optical and atomic force microscopy. The developed method represents a suitable approach for routine testing of food contamination.

Links

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

PASTUCHA, Matěj, Zdeněk FARKA, Tomáš JUŘÍK and Petr SKLÁDAL. Detection of Salmonella in Milk Using Enzymatic Precipitation Enhanced SPR Immunosensor. In CEITEC PhD Retreat II. 2017. ISBN 978-80-210-8550-3.

@proceedings{1379009,
   author = {Pastucha, Matěj and Farka, Zdeněk and Juřík, Tomáš and Skládal, Petr},
   booktitle = {CEITEC PhD Retreat II},
   keywords = {Surface Plasmon Resonance; Immunosensor; Salmonella; Enzymatic Precipitation; Food Safety},
   language = {eng},
   isbn = {978-80-210-8550-3},
   title = {Detection of Salmonella in Milk Using Enzymatic Precipitation Enhanced SPR Immunosensor},
   url = {https://www.ceitec.eu/ceitec-phd-retreat-ii-book-of-abstracts/f34779#page=99},
   year = {2017}
}

TY  - CONF
ID  - 1379009
AU  - Pastucha, Matěj - Farka, Zdeněk - Juřík, Tomáš - Skládal, Petr
PY  - 2017
TI  - Detection of Salmonella in Milk Using Enzymatic Precipitation Enhanced SPR Immunosensor
SN  - 9788021085503
KW  - Surface Plasmon Resonance
KW  - Immunosensor
KW  - Salmonella
KW  - Enzymatic Precipitation
KW  - Food Safety
UR  - https://www.ceitec.eu/ceitec-phd-retreat-ii-book-of-abstracts/f34779#page=99
N2  - Foodborne bacterial pathogens represent a serious threat for public health. Recent outbreaks have demonstrated the need for rapid and reliable screening techniques. As an alternative to the conventional methods, various biosensors have been developed. They offer fast, sensitive, specific, robust and cost effective analysis and have a potential for point-of-care application. In this work, we have developed a surface plasmon resonance (SPR) immunosensor for the detection of Salmonella Typhimurium in powdered milk. In addition to a conventional label free SPR assay, an enzymatic enhancement step was incorporated. The method did not require any pre enrichment, making the assay fast and simple. After Salmonella was bound by capture antibody immobilized on the sensor surface, solution of detection antibody labelled by horseradish peroxidase was injected. The bound enzymatic label in turn catalyzed the precipitation of 4-chloro-1-naphthol to insoluble benzo-4-chlorocyclohexadienone. The formation of precipitate on the sensor surface provided substantial signal enhancement. The precipitation-based assay format improved the sensitivity 40 times compared to the label-free format. The limit of detection of 10^2 CFU·mL-1 in buffer and 10^3 CFU·mL–1 in milk was achieved and the whole analysis time was below 60 min. The interaction of the bacteria with the sensor surface and the formation of the precipitate were studied in detail by optical and atomic force microscopy. The developed method represents a suitable approach for routine testing of food contamination.
ER  -

PASTUCHA, Matěj, Zdeněk FARKA, Tomáš JUŘÍK and Petr SKLÁDAL. Detection of Salmonella in Milk Using Enzymatic Precipitation Enhanced SPR Immunosensor. In \textit{CEITEC PhD Retreat II}. 2017. ISBN~978-80-210-8550-3.

Detailed Information on Publication Record