The Optimize Experimental Of Indirect Elisa Method To Detect F. Gigantica Of Cattle

Fasciolasis is a serious global parasitic zoonoses,caused by trematode that belong to Digenea,Fasciolidae,Fasciola,include F.hepatica and F.gigantica.The 2 trematodes were worldwide distribution,F.hepatica distribute in temperate zone and F.gigantica distribute in tropical and subtropical.Guangxi locate in subtropical,so F.gigantica is the dominant species.The annual economic loss caused by fasciolasis exceed 3.2 billion,and the number of infection people exceed 2.4 million,and 180 million of people have a risk of infection.So establish a time,labour and money saving method is especially important.In order to establish a method to detect multi-sample of F.gigantica,based on indirect ELISA,optimize the concentration coating buffer and coating time,kinds and concentration of blocking buffer and blocking time,concentration of serum and second-antibody,then detect the prevalence of F.gigantica in Guangxi with the methond.According to the results to establish a mutli-sample method.The concentration of serum and second-antibody are different between cow and buffalo.The positive serum was collected from buffalo that infected F.gigantica and the negative serum were collected from little cow and little buffalo that 1 week old.The optimize results by checkerboard method as follows:the best working concentration of ES is 1.6?L/mL for cow and buffalo.The best serum and second-antibody concentration of cow were 1:400 and 1:10000,and buffalo were 1:200 and 1:20000.Then decteted exceed 1000 serum samples collected from Naning,Liuzhou,Hezhou,Chongzuo and Baise with the method.According to the detection reulsts and OD to separate these samples,and act as the mateirals to establish mutli-sample indirect ELISA method for F.gigantica.Based on the optimized method to detect mix samples,and repeated to make sure the consistency and feasibility between the mutli-sample method and indirect ELISA method.The results showed that when the positive rate was 30%in the mix samples,the indirect mutli-sample ELISA method can dected the positive sample.