Design of a core-shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples

Ozalp, V. Cengiz
Öktem, Hüseyin Avni
We report an immuno-magnetic separation system developed by the immobilization of pathogen-specific antibodies on the core-shell magnetic beads. The magnetic beads were grafted with glycidylmethacrylate (GMA) using surface-initiated atom transfer radical polymerization (SI-ATRP). For immuno-magnetic separation (IMS) of target bacterial cells from others, antibodies for Escherichia coli and Salmonella enterica serovar Typhimurium cells were immobilized on the magnetic beads via glutaraldehyde coupling reaction. Our IMS system successfully separated Salmonella cells when the concentrations of target (i.e., Salmonella) and interfering (i.e., E. coli) cells were at the same level. Polymerase chain reaction (PCR) assays amplifying the rfb/rfbE region of the E. coli genome and a 647-bp fragment of the invA region of Salmonella were performed as the specific selection to accurately confirm the presence of E. coli and Salmonella, respectively. IMS and multiplex PCR methods can be used for specific and quantitative detection of pathogens from food samples. Thus, this study developed a reliable and direct system for rapid detection of Salmonella and E. coli in food samples. In addition, IMS method could be easily adapted to detect other pathogens by selecting the pertinent antibody.


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Demir, Ayhan Sıtkı; Talpur, Farah N.; Sopaci, S. Betul; Kohring, Gert-W; ÇELİK, AYHAN (Elsevier BV, 2011-04-10)
Rapid immobilization with the one-pot purification of galactitol dehydrogenase (GatDH) and formate dehydrogenase (FDH) is achieved by using iminodiacetic acid (IDA) with chelated Co2+ modified magnetic nanoparticles as a carrier. Lactate dehydrogenase (LDH) from recombinant Escherichia coli and FDH commencing Candida methylica were used as an auxiliary enzyme for the regeneration of NADH/NAD(+) with a representative synthesis of (S)-1,2-propanediol and L-tagatose starting from hydroxyacetone and galactitol....
Evaluation of structural changes induced by high hydrostatic pressure in Leuconostoc mesenteroides
Kaletunc, G; Lee, J; Alpas, Hami; Bozoglu, F (American Society for Microbiology, 2004-02-01)
Scanning electron microcopy (SEM), transmission electron microscopy (TEM), and differential scanning calorimetry (DSC) were used to evaluate structural changes in Leuconostoc mesenteroides cells as a function of high-hydrostatic-pressure treatment. This bacterium usually grows in chains of cells, which were increasingly dechained at elevated pressures. High-pressure treatments at 250 and 500 MPa also caused changes in the external surface and internal structure of cells. Dechaining and blister formation on ...
Analysis of acyl CoA ester intermediates of the mevalonate pathway in Saccharomyces cerevisiae.
Şeker, Tamay; Nielsen, J (Springer Science and Business Media LLC, 2005-04-01)
The mevalonate pathway plays an important role in providing the cell with a number of essential precursors for the synthesis of biomass constituents. With respect to their chemical structure, the metabolites of this pathway can be divided into two groups: acyl esters [acetoacetyl CoA, acetyl CoA, hydroxymethylglutaryl (HMG) CoA] and phosphorylated metabolites (isopentenyl pyrophosphate, dimethylallyl pyrophosphate, geranyl pyrophosphate, farnesyl pyrophosphate). In this study, we developed a method for the ...
Design of intelligent nanoparticles for use in controlled release
Bayyurt, Banu; Hasırcı, Vasıf Nejat; Department of Biotechnology (2009)
The aim of this project was to design an intelligent controlled release system based on thermoresponsive nanoparticles for cancer therapy and to evaluate the efficiencies of these systems with in vitro cell culture. Poly(Nisopropylacrylamide), an important thermoresponsive polymer, was selected for this study to prepare the responsive nanoparticles. This polymer has an lower critical solution temperature (LCST) of 32 oC, below which it is hydrophilic and above this temperature, it shows hydrophobic behavior...
MCCARTHY, MJ; PEREZ, E; OZILGEN, M (Wiley, 1991-11-01)
The use of magnetic resonance imaging (MRI) spectroscopy as a real time probe to noninvasively obtain transient moisture profiles during food dehydration is demonstrated. A laboratory-scale wind tunnel drier was constructed. A 2 x 5 x 5 cm3 apple slab was inserted in the probative zone and one-dimensional drying was effected at quasiisothermal conditions. Spatial resolution of 0.60 mm was obtained. The transient moisture profiles were simulated with a simple mass transfer model. Effective diffusivity of wat...
Citation Formats
V. C. Ozalp, G. BAYRAMOĞLU, M. Y. ARICA, and H. A. Öktem, “Design of a core-shell type immuno-magnetic separation system and multiplex PCR for rapid detection of pathogens from food samples,” APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, pp. 9541–9551, 2013, Accessed: 00, 2020. [Online]. Available: