Establishment and application of real-time PCR for the detection of Toxoplasma gondii

The specific primers were designed targeting the 529 bp fragment of Toxoplasma gondii RH strain. A 212 bp specific andconsensus fragment was amplified and cloned into p GEM-T Easy vector. Standard curve was generated by using serial dilutions of re-combinant plasmid,showing that the correlation coefficient was 0.997. The mean interassay coefficient of variation was 2.30%. The detectionof T.gondii YZ-1 strain and YZ-2 strain were positive. The detection of Sarcocystis fusiformis,Isospora canis and Eimeria tenella werenegative. The twenty-four dead pigs from animal hospital were deteced with this method and conventional PCR. The positive rate was 70.8%as exmined by real-time fluorescent quantitative PCR,and 66.7% by conventional PCR. However,as to the detection results of 69 tissuesamples from 24 dead pigs,the positive rate(50.7%)by real-time fluorescent quantitative PCR was apparently higher than that(34.8%)by conventional PCR.