Abstract
Pathogen separation is of great significance for precise detection and prevention of disease outbreaks. For the first time, protein A conjugated with chitosan-coated iron oxide nanoparticles was prepared for pathogen separation at low concentrations from liquid samples. Vibrio cholerae O1 (VO1) bacteria were used for testing the effectiveness of this conjugate. Transmission electron microscopy (TEM) was used to confirm the presence of captured VO1. The results showed that, after binding with a specific antibody, the conjugate allows separation of VO1 bacteria from water samples at a concentration as low as 10 cfu mL(-1). Moreover, the conjugate can be used in parallel with conventional or modern diagnostic tests for quick and accurate detection of pathogens.
Highlights
The world has witnessed an increased number of emerging and reemerging infectious disease outbreaks caused by viruses and bacteria, such as ebola, severe acute respiratory syndrome-associated coronavirus (SARS-CoV), avian in uenza (A/H5N1, A/H7N9), foot–hand–mouth diseases, measles, diarrhea and cholera
The uorescent labeling technique is preferred for the detection of V. cholerae in environmental water samples, but ltering is necessary to concentrate the bacteria from a large volume of water
The obtained system is highly stable due to steric repulsion of the nanoparticles provided by the polymer backbone and the surface charge obtained from the amine groups
Summary
The world has witnessed an increased number of emerging and reemerging infectious disease outbreaks caused by viruses and bacteria, such as ebola, severe acute respiratory syndrome-associated coronavirus (SARS-CoV), avian in uenza (A/H5N1, A/H7N9), foot–hand–mouth diseases, measles, diarrhea and cholera. A polysaccharide obtained from the N-deacetylation of chitin, is one of the most abundant polysaccharides in nature It has been used for a wide range of biological applications, thanks to its biocompatibility and biodegradability.[8] Its use for the functionalisation of nanoparticles allows the incorporation of amine groups on the surface of the nanoparticles, and adds a positive charge to the system, which has been shown to be preferred by some cell lines.[9] On the other hand, protein A is well known as a molecule which binds with high affinity to the Fc region of IgG antibodies and orientates the binding sites of the antibody molecule outwards from the surface of nanoparticles for capturing antigens.[10]. The V. cholerae bacteria could be magnetically separated and collected by binding to iron oxide nanoparticles
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