Immune Colloidal Gold Test Strip For The Development Utend Rapid Detection Of Goat Toxoplasmosis

Toxoplasmosis an important zoonotic parasitic diseases caused by Toxoplasma gondii. The disease is worldwide distribution, infection with a wide range of human and animal safety risk. The current vaccine to prevent toxoplasmosis is mainly genetically engineered vaccine, only applies to animals, immunity, and vaccine safety and effectiveness can not be guaranteed, for the people of Toxoplasma gondii vaccine still in animal testing stage, so diagnosis of Toxoplasma gondii Research is very important. Current diagnosis of toxoplasmosis is divided into serological diagnosis, diagnostic immunology and molecuLar biology diagnosis, serological diagnosis of specific weak, prone to misdiagnosis; molecuLar biology diagnostics complex and time-consuming, so the immune diagnosis of a commonly used testing methods. Immunogold detection technique the immune diagnosis method, with its simple, sensitive and specific, strong stability, does not require special equipment and reagents result, the advantages of intuitive judgments, is widely used in pathogenic microorganisms, toxins, drug residues, etc. detection.This test uses recombinant Toxoplasma gondii Surface Protein SAG2 (P22) as an antigen, using sodium citrate reduction method to colloidal gold, and Staphylococcal protein A (SPA) combine to form probe, probe purified by ultracentrifugation on Preparation conditions of strip filter assembly rapid detection of goat immune colloidal gold test strip, and strips of sensitivity, specificity, and reliability analysis.The test gene was amplified from T. gondii SAG2 construct pGEX-4T-1-SAG2 recombinant plasmid, E. coli and gel purified, recombinant SAG2 high purity protein. Screening of the prepared strips obtained optimum conditions:20nm particle size of colloidal gold in pH6.4 formed under the conditions of tags 25ug/mLSPA combination of the most solid probe to sodium borate diluted probe to BSA+NaN3+polyethylene glycol 10 000 as a probe stabilizer mixture to Sartorius CN140-based nitrocellulose membrane as a carrier, recombinant SAG2 l.Omg/mL detect protein-coated wire, horse anti-goat IgG to 0.9mg/mL coated control line and assembly of immuno-colloidal gold test strip for detection of the best. Strip for detection of Toxoplasma-positive serum sensitivity of the 1:5120, and goat coccidia positive serum, goat brucellosis, S. japonicum sera no cross reaction.