Establishment of a Colloidal Gold Immunochromatographic Assay for Detecting Fasciola hepatica Infection in Sheep Based on Fh010935 Protein
A colloidal gold immunochromatographic assay for detecting Fasciola hepatica infection in sheep was established using recombinant protein Fh010935.Firstly,colloidal gold solution was prepared,pH value of the colloidal gold solution and SPA labeling amount were optimized,and the gold labeled protein was identified by transmission electron microscopy.Then coating concentration of(T)line recombinant protein Fh0 10935 and control(C)line anti SPA antibody were optimized and detected,and colloidal gold immunochromatographic strips were prepared.Finally,sensitivity,speci-ficity,repeatability and preservation period of the strips were evaluated,and sheep serum samples from Inner Mongolia were tested.The results showed that the optimal concentration of K2CO3 was 0.4 mmol/L,and the optimal labeling amount of SPA was 6 μg/mL,the optimal coating concentration of T line was 0.25 mg/mL of Fh010935 recombinant protein,and the optimal coating concentration of C line was 8 times of diluted anti SPA antibody.The performance test showed that the minimum se-rum dilution of this method was 1:160,and it had no immune cross reaction with the sera infected with Haemonchus contortus,Dicrocoelium lanceatum and Neospora caninum,and had good repeatabil-ity.The storage period at 4 ℃ and at room temperature reached 180 days.The positive rate of 64 sheep serum samples in Inner Mongolia was 14.1％.The colloidal gold immunochromatographic assay for detecting F.hepatica infection in sheep established with the recombinant protein Fh010935 as the coating antigen has good sensitivity,specificity and repeatability,and a long storage period.It can be used for rapid detection and epidemiological investigation of F.hepatica infection in sheep.
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