The Research About GICA And ELISA Of M.Suis

Eperythrozoonosis is a zoonosis caused by Eperythrozoon suis.It has host-specificity, often causes fever, hemolysis, jaundice and other symptoms. This disease is universaland recessive infection. The clinical symptoms are similar to many other diseases, what is m ore, the disease is easy to misdiagnosis and could bring some hidden troubles to Pig industry. Therefore, diagnosis of eperythrozoonosis is the key to prevent and control it. However, the current detecting method is still lacking of a practical and convenient way for veterinarians to mass detection and to do large scale survey. The advantage of ELI SA and GICA methods, which are simple, fast, stability, high sensitivity, makes ELISA and GICA become the main method for animal disease detection. Aditionally, the GICA methods to detect the antibody against M.suis was seldom reported.In this paper,eperythr ozoon antigen was cultured in our laboratory, and optimized IHA method fordetection of M.suis. Then the indirect ELISA and GICA method were established to detect the antib-ody against M.suis.In this paper,three methods were compared, to seek the best method.This study is divided into four parts:(1) I used laboratory cultured porcine M.suis immuning pigs, and obtained positive serum. The titer was1:512.(2) Optimization of IHA test method, concentration of M.suis2mg/mL sensitized red cell45min at37℃, then402clinical samples were detected.(3) This study established an indirectable ELISA method. The optimum conditions were that quality of antigen coated is80ug per hole,0.5%BSA coated overnight at4℃; closed one hours at37℃plus closed one hou rs at4℃; serum dilution of1:40reacted1.5hours;HRP-IgG antibody diluted1:1000rol e of45min;finally, TMB37℃react20min. ELISA results showed as follows, when OD450≥0.195be positive; when OD450<0.195found negative.(4) The study preparation of24.5nm gold particles, the preparation of gold probe Rabbit anti pig IgG and colloidal gold combined, antigen of2mg/mL and positive serum were used as the detection line and control line coated NC membrane; used3%BSA+0.5%Tween-20closed NC membra ne one hour at37℃;gold standard pad1%BSA+0.5%Tween-20+5%sucrose+0.5%PEG5000pretreatment at37℃drying; The gold pad used1%BSA+0.5%Tween-20+5% sucrose+0.5%PEG5000pretreatment and drying of37℃.402serum samples in22swine-farms in Hunan province were detected by IHA, ELISA, GICA,and the positive rates were53.73%,61.69%,53%,respectively.Comparison among the result detected by IHA、ELISA、GICA, it revealed a significant correlation (P>0.05). The conclusion could be draw that ELSIA and GICA could be used for the d iagnosis and epidemiological survey of M.suis in basic laboratory, but GICA have better convenience,sensitivity,efficiency, and stability,which is more suitable for mass detection and population survey.