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Diazonium-based impedimetric aptasensor for the rapid label-free detection of Salmonella typhimurium in food sample
Linköping University, Department of Physics, Chemistry and Biology, Biosensors and Bioelectronics. Linköping University, Faculty of Science & Engineering. University of Mazandaran, Iran.
University of Mazandaran, Iran.
Linköping University, Department of Physics, Chemistry and Biology. Linköping University, Faculty of Science & Engineering.
Linköping University, Department of Physics, Chemistry and Biology, Biosensors and Bioelectronics. Linköping University, Faculty of Science & Engineering.ORCID iD: 0000-0002-1815-9699
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2016 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 80, p. 566-573Article in journal (Refereed) Published
Resource type
Text
Abstract [en]

Fast and accurate detection of microorganisms is of key importance in clinical analysis and in food and water quality monitoring. Salmonella typhimurium is responsible for about a third of all cases of food borne diseases and consequently, its fast detection is of great importance for ensuring the safety of foodstuffs. We report the development of a label-free impedimetric aptamer-based biosensor for S. typhimurium detection. The aptamer biosensor was fabricated by grafting a diazonium-supporting layer onto screen printed carbon electrodes (SPEs), via electrochemical or chemical approaches, followed by chemical immobilisation of aminated-aptamer. FTIR-ATR, contact angle and electrochemical measurements were used to monitor the fabrication process. Results showed that electrochemical immobilisation of the diazonium-grafting layer allowed the formation of a denser aptamer layer, which resulted in higher sensitivity. The developed aptamer-biosensor responded linearly, on a logarithm scale, over the concentration range 1 x 10(1) to 1 x 10(8) CFU mL(-1), with a limit of quantification (LOQ) of 1 x 10(1) CFU mL(-1) and a limit of detection (LOD) of 6 CFU mL(-1). Selectivity studies showed that the aptamer biosensor could discriminate S. typhimurium from 6 other model bacteria strains. Finally, recovery studies demonstrated its suitability for the detection of S. typhimurium in spiked (1 x 10(2), 1 x 10(4) and 1 x 10(6) CFU mL(-1)) apple juice samples. (C) 2016 Elsevier B.V. All rights reserved.

Place, publisher, year, edition, pages
ELSEVIER ADVANCED TECHNOLOGY , 2016. Vol. 80, p. 566-573
Keywords [en]
Diazonium grafting; Aptamer; S. typhimurium; Label-free detection; Electrochemical impedance spectroscopy; Food analysis
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:liu:diva-127249DOI: 10.1016/j.bios.2016.02.024ISI: 000372558500079PubMedID: 26894987OAI: oai:DiVA.org:liu-127249DiVA, id: diva2:921644
Note

Funding Agencies|Vetenskapsradet (Pathoscreen project; Swedish Research Link) [D0675001]; Ministry of Science Research and Technology of Iran

Available from: 2016-04-20 Created: 2016-04-19 Last updated: 2019-10-09Bibliographically approved
In thesis
1. Functionalised surfaces for bacterial discrimination
Open this publication in new window or tab >>Functionalised surfaces for bacterial discrimination
2016 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Bacterial detection and identification is a critical step in many arenas, including food and water safety, clinical diagnostics, bioprocess control and biosecurity. Social hygiene has a direct correlation with the strict control of microorganisms in these fields. The worldwide cases of bacterial infectious disease is assessed to be 1-2 billion annually, and these have a massive negative effect on the global economy. Although many precise techniques are currently available, a huge mortality and morbidity related to bacterial infection disease continues to be reported annually due to misdiagnosis or delay in diagnosis. Increasing efficiency and reliability of pathogen detection methods will potentially improve social health and protect society against pathogenic diseases.

The development of culture media for selective isolation and differentiation of bacteria started in the late 19th century. Immunological assays and then genotyping techniques were developed in 20th century, in addition to many less commonly used techniques for bacterial detection. Each of the currently used methods has its advantages and weaknesses in terms of speed, cost and accuracy. Much effort has recently been devoted to developing biosensors for bacterial detection for simpler and more rapid use.

This thesis is focused on functionalised surfaces for the development of biosensors for bacterial discrimination and detection, and is divided in three subsections. First, we explored a new approach for bacterial discrimination based on pattern recognition. Traditional culturing methods discriminate bacteria based on their metabolic activity pattern. Taking inspiration from the extensive body of work that reports the use of electronic-noses to differentiate bacteria based on the volatiles patterns they produce, we explored the possibility of bacteria differentiation based on adhesion patterns. By altering the electropolymerisation conditions, the physicalchemical surface properties of polypyrrole (PPy) can be tuned to fabricate a range of dissimilar surfaces. The adhesion of different bacteria on a series of polymers was measured. Data analysis of the adhesion patterns proved that bacteria can be discriminated by examining their adhesion to dissimilar surfaces. Next, we developed a new functionalisation of PPy by doping PPy with 4-N-Pentylphenylboronic Acid and investigated the modulation of bacteria binding to those surfaces. In this second section, a new electropolymerisation technique for whole-cell imprinting was developed based on different functional monomers. 3-Aminophenyl boronic acid was shown to be a good monomer to produce whole-cell imprinted polymers (CIP) with high affinity for bacterial cells with improved releasing ability. Finally, in the third section aptamers, which are promising synthetic recognition elements, were explored for bacterial detection testing. A specific aptamer was used to fabricate of a prototype of label-free aptasensor for bacterial detection. Also, the SELEX process was used to produce a pool of aptamers, or “polyclonal” aptamers, which targeted a group of bacteria species. Using polyclonal aptamers as a recognition element enables biosensors to enhance their resolution to detect broader types of bacterial species using a single serological-like test.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2016. p. 55
Series
Linköping Studies in Science and Technology. Dissertations, ISSN 0345-7524 ; 1770
Keywords
Biosensorer, Bakterier, Mönsterigenkänning
National Category
Bio Materials
Identifiers
urn:nbn:se:liu:diva-160803 (URN)9789176857519 (ISBN)
Public defence
2016-06-17, Nobel, B-huset, Campus Valla, Linköping, 10:15 (English)
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Supervisors
Available from: 2019-10-09 Created: 2019-10-09 Last updated: 2019-10-09Bibliographically approved

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Golabi, MohsenTurner, AnthonyBeni, Valerio

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