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The Staphylococcus aureus Exotoxin Recognition Using a Sensor Designed by Nanosilica and SEA genotyping by Multiplex PCR

H. Ahari, B. Akbari-Adreghani, V. Razavilar, A. Motallebi, S. Moradi, A.A. Anvar
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Abstract

Considering the ever increasing population and industrialization of the developmental trend of human life, we are no longer able to detect the toxins produced in food products using the traditional techniques. This is due to the fact that the isolation time for food products is not cost-effective, and even in most of the cases, the precision of practical techniques like bacterial cultivation and other techniques suffers from operator errors, or the errors of the mixtures used. Hence, with the advent of nanotechnology, the design of selective and smart sensors has turned into one of the greatest industrial revelations of the quality control of food products that, in few minutes time and with a very high precision, can identify the volume and toxicity of the bacteria. In this research, based on the bacterial antibody's connection to nanoparticles, a sensor was used. In this part of the research, as the basis for absorption for the recognition of bacterial toxin, medium sized silica nanoparticles of 10 nm in the form of solid powder were utilized with Notrino brand. Then the suspension produced from the agent-linked nanosilica, which was connected to the bacterial antibody, was positioned near the samples of distilled water, which were contaminated with Staphylococcus aureus bacterial toxin with the density of  10-3 molar, so that in case any toxin exists in the sample, a connection between the toxin antigen and the antibody would be formed. Finally, the light absorption related to the connection of antigen to the particle-attached antibody was measured using spectrophotometry. The 23S rRNA gene that is conserved in all Staphylococcus spp. was used as the control. The accuracy of the test was monitored by using the serial dilution (l0-6) of overnight cell culture of Staphylococcus spp. bacteria (OD600: 0.02 = 107 cell). It showed that the sensitivity of PCR is 10 bacteria per ml of cells within few hours. The results indicated that the sensor detects up to 10-4molar density. Additionally, the sensitivity of the sensor was examined after 60 days; by the 56 day, it had confirmatory results, which started to decrease after this time. Comparison of practical nanobiosensory method with the conventional methods including culture and bio-technology methods (such as polymerase chain reaction) confirmed its accuracy, sensitiveness and uniqueness.  It also reduces the time from hours to 30 minutes.

Keywords

Nanosensor, Staphylococcus aureus, Exotoxin, Multiplex PCR

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DOI: https://doi.org/10.22037/afb.v1i1.7126

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