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Preparation and Detection of Egg Yolk Antibody Against Streptococcus Bacteria in Pharynx and Throat

Author: LiuDuan
Tutor: Zhang;ZhaoSuQing
School: Guangdong University of Technology
Course: Applied Chemistry
Keywords: Streptococcus pneumoniae MG hemolytic streptococcus Yolk antibody Titer Preparation Detect Antibacterial Test
CLC: R392
Type: Master's thesis
Year: 2011
Downloads: 37
Quote: 0
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Abstract


Mouth and throat disease is a common human diseases, mainly caused due to bacterial infection, determined after research, the main infection bacteria Streptococcus, Staphylococcus, and Streptococcus pneumoniae and other. Mouth and throat diseases will not immediately cause human death, but because of its diversity of diseases, easy to relapse, it is difficult to cure, and bring a lot of inconvenience and trouble it caused many complications to the life and work of human. Therefore, the attention has been on the treatment of diseases of the mouth and throat. Human anti-bacterial and virus is mainly dependent on antibiotics, but due to antibiotic treatment will produce many adverse consequences, including physical function coordination mechanism disorders of side effects, bacteria, virus resistance increases the odds. In recent years, starting from the point of view of the immunological study found that using the IgY technology can produce antibodies against drug-resistant treatable disease caused by bacterial or viral infections, has security no residue, do not become resistant, inexpensive The advantage of a broad application prospects antibiotic alternatives. We prepared anti-Streptococcus pneumoniae, MG hemolytic streptococcus specific yolk antibodies, then by SDS-PAGE gel electrophoresis, Coomassie blue staining, antibacterial test, enzyme-linked immunosorbent assay, cross-reactivity test its biological activity were identified, and IgY thermal stability, pH stability and stability of digestive enzymes have been studied. The results showed that IgY purity acetic acid - sodium acetate buffer, PEG 6000 and ammonium sulfate solution three-step purification to get up to about 95%; indirect enzyme-linked immunosorbent assay (ELISA) showed that hens for the first time the immune 10d, yolk ie antibody producing, initial immunization, 20 d aftereffect price gradually increased, high titer appear in the 40-d-80 d after the initial immunization, the titer of specific egg yolk antibody against Streptococcus pneumoniae can be up to 1:5120, anti-MG hemolytic streptococcus specific egg yolk antibody titer can reach up to 1:2560; vitro the antibacterial experiments found specific IgY against Streptococcus pneumoniae, MG hemolytic streptococcus can significantly inhibit the inhibitory rate can reach 90%; cross Reaction experiment prepared specific IgY is capable of specifically binding to its antigen bacteria. In summary, this study successfully prepared by a high-content, high purity and specificity of the anti-Streptococcus pneumoniae, anti-MG the hemolytic streptococcus specificity IgY, bacteriostatic in vitro experimental studies have shown its ability to effectively combined with bacteria and their antigens bacteria have a good inhibitory effect, laid the foundation for the prevention and control applications in pharyngitis specific IgY.

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