Universal enzyme-linked immunosorbent assays (ELISA) and utility in the detection of antibodies against Salmonella spp. in several animal species

Abstract

The aims of this research are confirming the feasibility of hybrid immunoglobulin-binding reagents, its used in ELISAs for IgG/IgY detection and detecting specific antibodies against an infectious microorganism (Salmonella spp) in various animal species using a universal diagnostic ELISA. Hybrid immunoglobulin-binding bacterial proteins (IBP) including recombinant protein LA, recombinant protein LG, and recombinant protein AG have been produced for improvement of their binding affinity to a much larger number of immunoglobulins. This hybrid bacterial protein thus represents a powerful tool for binding, detection, and purification of immunoglobulins and their fragments. However, SpLA-LG-peroxidase and SpLAG-anti-IgY-peroxidase were produced by the periodate method. They have shown to be effective reagents. Their binding affinity to immunoglobulins surpasses previous hybrid IgG-binding proteins reported, including the most known SpAG, SpLA and SpLG. The IgY fraction was isolated from the egg yolks of a variety of birds including species of chicken, bantam hen, guinea hen, quail, goose, duck, wild and domestic pigeon, parakeet, cattle egret, pheasant, and ostrich. The IgY fraction was isolated by the chloroform-polyethylene glycol (PEG) method. An ELISA for anti-Salmonella spp antibodies was employed with some modifications to determine the presence of antibodies in humans, laying hens, geese, quails, and pigeons. Salmonella are motile, flagellated rod-shaped zoonotic pathogens which may survive with or without oxygen. They belong to the family Enterobacteriaceae and is implicated with typhoid fever and food-borne illnesses. This pathogen is associated with several diseases, which may become fatal and negatively impact the health of individuals and various economies globally. The poultry industry is most impacted and vulnerable to the onslaught of this pernicious microbe.

Universal Enzyme-Linked Immunosorbent Assays (ELISA) and Utility in the Detection of Antibodies against Salmonella spp. in Several Animal Species

“…In addition, they have been used in the diagnosis of zoonotic infections in tens of zoo animals worldwide. Thus, this hybrid bacterial protein represents a powerful tool for the binding, detection, and purification of immunoglobulins and their fragments [1].…”

“…1 shows SDS-PAGE, which depicts the creation of fusion proteins to detect antibodies. Several hybrid proteins, which have been extensively studied, can be visualized by protein electrophoresis [1].…”

“…The lipopolysaccharide somatic O antigen, flagellar H, and virulent V antigenic structures determine the serotype designate Salmonella species [1].…”

“…Salmonella resists the acidic conditions imposed by gastric juice and amino acids in the stomach. Incubation may last for six to twenty-four hours [1].…”

“…One may experience stomach cramps, vomiting, nausea, diarrhea, ulcers, and elevated body temperature. Systemic breach of blood vessels and entry of bacteria into the bloodstream may be fatal [1]. Salmonellosis persists for up to 4-7 days.…”