The Establishment Of Tissue PCR Diagnosis Method For Cattle Neospora Caninum

Neosporiasis is a kind of protozoal disease caused by neospora caniumparasitizes in various kindsof livestock such as dog, cattle, horse, andsheep. This disease could bring the cow abortion, even if the foetus couldchildbear, the foetus constitution would be weaker, also. Penez consideredthat Neosporiasis is the main reason that caused cosmopolitan cowabortion, its mode of transmission maybe like T. gondii, some of theminfected tachyzoite by devouring oocyst that was in the long run host, or by mouth or nose, but the vertical transmission was main way. Thus, Neosporiasis is a kind of fairly severe parasitosis to harm the husbandryand cow cultivation.This experiment took gerbil as the animal model, and succeed inextracting Neospora caninum DNA from artificial infection gerbil tissue.Then, we designed a couple specificity primer based on the Nc5 gene orderreported on the Genbank, having the PCR amplification, obtaining a singleband gene fragment size 338bp, initially establishing the PCR detectiontechnique of cattle Neosporiasis, at the same time, having the specificityexperiment and the sensibility experiment The experiment results showedthat this sequences did not amplificate DNA of cryptozoite, T. gondii andcattle Theileria sergenti, when the DNA contents of neospora caninum was1.8 fg/ul, we Could amplificate the clear band.We had the gene cloning in the pMD18-T vector, by PCR and enzymecutting identificated the recombinant plasmid, then having the sequencinganalysis, the results showed that the clonal gene was neospora caninum specificity gene.Having the IFAT detection to 10 aborted cows from wangqing, helong, longjing and chaoyangchuan, choosing 1 aborted fetus that was positivein IFAT experiments to have PCR diagosis, the results showed that it waspositive.