Cultivation Of Fasciola Gigantica Metacercariae And Establishment And Application Of Single Antibody Sandwich Elisa Method

Fasciola gigantica is a kind of distribution in China’s south and southeast Asia area common parasites,many parasitize animals such as cows and goats liver bile duct,which cause serious damage to the animal’s normal growth and development.Fasciola gigantica is a serious zoonotic parasitic disease,in 2011 China’s Yunnan Province struck Binchuan suffered large crowd disease,due to the lack of effective diagnostic monitoring method,causing major outbreaks,a total of more than 20 people infected.The disease diagnosis,treatment and prevention of research work is still relatively weak.Especially still lack of practical and effective diagnostic and the immune preventive methods,thus establishing a specific and sensitive method of early diagnosis is the necessary.This study using artificial breeding Galba pervia,the intermediate host of artificial cultivating Fasciola gigantica trematode eggs hatch miracidium,with Fasciola gigantica capsule infected Galba pervia to be successful,the temperature 30 ℃,12 d hatched miracidium,and mice as experimental animals for the artificial infection test,the results show that Fasciola gigantica larvae survival in mice development 7 to 8 weeks,and can cause more regular pathological changes.Five hybridoma cell lines,including 6D3,6B4,7D2,7D1 and 7D4,eventually matched using monoclonal antibodies 7D2 and 7D1.The optimal coating concentration of monoclonal antibody was 1:6400,the optimal working dilution of HRP-labelled 7D1 Ig G was1:10000,and the reaction time of serum was 1h,the reaction time of HRP-labelled 7D1 was 1h,the reaction time of substrate was 25min.The ELISA had nocross-reaction with Eurytrema pancreaticum,,Paramphistomum,Trypanosoma evansi,Toxoplasma gondii,this method can be diagnosed early Fasciola gigantica disease infection status.This study also used trichlorobenzene azole,albendazole treatment,praziquantel artificial feeding mice infected with the Fasciola gigantica trematode capsule larva.The results showed that three chlorobenzene of good treatment effect,autopsy observation,in the liver of the abdominal bug rate is low,no obvious pathological changes.This work could provide the theoretical basis for the reasonable application of single antibody sandwich ELISA method,and it will basis for further research of Fasciola gigantica.