Gold nanoparticles (AuNPs) of 15-20 nm size range have attracted attention for producing smart sensing devices as diagnostic tools in biomedical sciences. Citrate capped AuNPs are negatively charged, which can be exploited for electrostatic interactions with some positively charged biomolecules like antibodies. In this paper we describe a method for the low cost synthesis of gold nanoparticles using sodium citrate (Na 3 Ct) reduction in chloroauric acid (HAuCl 4 .3H 2 O) by microwave heating (diameter about 13-15 nm). Gold nanoparticles were functionalized with surface activation by 3-mercaptopropionic acid for attaching antibody. These nanoparticles were then reacted with anti-E. coli O157:H7, using N-hydroxy succinimide (NHS) and carbondimide hydrochloride (EDC) coupling chemistry. The product was characterized with UV-visible spectroscopy, Fourier transform infrared (FTIR) spectroscopy and zeta potential. In addition, the binding of antibody-gold nanoparticles conjugates to E. coli O157:H7 was demonstrated using transmission electron microscopy (TEM).
Although Escherichia coli (E. coli) is a commensalism organism in the intestine of humans and warm-blooded animals, it can be toxic at higher density and causes diseases, especially the highly toxic E. coli O157:H7. In this paper a quartz crystal microbalance (QCM) biosensor was developed for the detection of E. coli O157:H7 bacteria. The anti-E. coli O157:H7 antibodies were immobilized on a self-assembly monolayer (SAM) modified 5 MHz AT-cut quartz crystal resonator. The SAMs were activated with 16-mercaptopropanoic acid, in the presence of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and ester N-hydroxysuccinimide (NHS). The result of changing frequency due to the adsorption of E. coli O157:H7 was measured by the QCM biosensor system designed and fabricated by ICDREC-VNUHCM. This system gave good results in the range of 102–107 CFU mL−1 E. coli O157:H7. The time of bacteria E. coli O157:H7 detection in the sample was about 50 m. Besides, QCM biosensor from SAM method was comparable to protein A method-based piezoelectric immunosensor in terms of the amount of immobilized antibodies and detection sensitivity.
In the present work a low cost technique for preparation of gold nanoparticles (AuNPs) using microwave heating was developed. The effect of different elements (precursor reagents, irradiation time, and microwave radiation power) on the final morphology of AuNPs obtained through the microwave assisted technique has been investigated. The characterization of the samples has been carried out by transmission electron microscopy, UV-vis absorption spectroscopy, Fourier transform infrared spectroscopy, and powder x-ray diffraction. The results showed that to some extent the above-mentioned characterizations influenced the size of synthetized nanoparticles and application of microwave heating has many advantages such as low cost, rapid preparation and highly uniform particles. As an application in quartz crystal microbalance (QCM) immunosensor, AuNPs are conjugated with the Escherichia coli (E.coli) O157:H7 antibodies for signal amplification to detect E.coli O157:H7 bacteria residual in QCM system.
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