Objective To establish a rabbit model of Eimeria stephensi infection and a nested PCR diagnostic method.
Methods The rabbit E. stiedai model was identified by autopsy, microscopic observation, blood biochemistry and histopathological examination. By collecting oocysts, extract E We designed specific primers to detect the DNA of stiedai and established a nested PCR method for E. stiedai.
Results Clinical autopsy showed that the abdominal circumference of rabbits was significantly increased. Anatomically, the liver was enlarged, the surface and substance of the liver were covered with white and yellowish nodules, the gallbladder and bile duct were enlarged, and the bile was yellowish. Microscopic observation: egg size is (31 72~38.43) μ m × (18.10~22.69) μ um。 Blood biochemical examination: globulin (GLOB) index was high, creatinine (CREA) and a magnetic acid cutoff (ALKP) index were low. Other test indexes are within the range of reference value. Histopathological examination: liver tissue and
A large amount of pink E Stiedai eggs. Nested PCR: the minimum detection limit was 1 oocyst DNA sample and 1.14 x 103 copy number plasmids. No bands were found in the negative control and blank control, and the coefficient of variability of the repeatability experiment was<5%.
Conclusion The rabbit E The nested PCR diagnostic method established by stiedai infection model can amplify E The specific fragment of stiedai has strong sensitivity and good repeatability.