Preparation Of Immunity Colloidal Gold Test Strip For Detecting Fasciola Gigantica Disease

Fasciolasis,caused by Fasciola gigantica,is distributed all over the world.The ruminants such as sheep,cattle,and buffalo with the Fasciolasis can induce acute,chronic,or sub-clinical symptoms like hepatitis and cholangitis.Sometimes Fasciolasis causes considerable economic losses in livestock and threatens the human's health.The prevention and treatment of this disease is dependent on the effective and efficient detection method.In this study,a new diagnostic method for detecting Fasciola gigantica was established by immunological and colloidal gold chromatographic techniques.For preparing the test strip,the excretory-secretary antigen(ES Ag)of Fasciola gigantica and the monoclonal antibodies of hybridoma cell strains 7D1 and 7D4 against excretory-secretary antigen of Fasciola gigantica were prepared and purified.Meanwhile,the colloidal gold particle was prepared by the sodium cirrate reduction method,then the affinity purified 7D1 monoclone antibodies was labeled with the 28 nm colloidal gold particles.The 7D4 monoclonal antibodies and the goat anti-mouse IgG antibody using as secondary antibodies were coated on the surface of nitrocellulose filter membrane(NC)as the test line(T line)and control line(C line)respectively.By optimizing the experiment conditions,the test strip was assembled with the materials above.The test results showed the prepared strip had a detecting prescribed minimum of 0.94?g/mL of Fasciola gigantica excretory-secretary antigen(ES Ag),and it has no any cross reaction with the antigens of Eurytrema pancreaticum,paramphistome,Chlamydia,Toxoplasma gondii,and Trypanosoma evansi.19 positive and 35 negative fecal samples were tested by using the strip and ELISA and then the results indicated the coincidence rate evaluated in depth were 87.04%and 94.44%in the detection respectively.334 fresh clinical samples of water buffaloes' feces in Guangxi were collected and were detected by using the prepared strip and the positive rate was 38.62%.These results indicated that this strip was a sensitive,specific and reliable method for detecting Fasciola gigantica ES antigens.The establishment of test strip for detecting Fasciola gigantica disease has provided a new method for the diagnosis and epidemiological investigation of Fasciolasis.