Development and Application of Nested PCR Assay for Detection of Toxoplasma gondii Infection in Dogs
Toxoplasmosis is a global zoonosis,which requires a rapid and sensitive method for the detection of canine infection.Nested PCR primers were designed according to the repetitive 529 bp DNA fragment of Toxoplasma gondii in GenBank.The nested PCR detection method was then developed and optimized for its reaction conditions.The results of specificity test showed that this method had no amplification of Cattle Babesia,Theileria annulata and Neospora.The sensitivity test results showed that it was 100 times higher than the national standard PCR and the method detected the lowest T.gondii gene of 0.134 pg.Testing of tissue samples from dogs infected with T.gondii showed good detection by this nested PCR.Taken together,the nested PCR method developed here had strong specificity,high sensitivity and good repeatability,which provided technical support for the detection of T.gondii infection and prevention and control of the disease.
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