Genotyping of Toxoplasma gondii using three important genes B1, GRA6 and Rep-529

Toxoplasma gondii is one of the most pathogenic zoonoses parasite can cause diseases both in humans and different species of birds and mammals. Immunocompromised patients and pregnant women are the most sensitive groups that can be seriously affected by the disease, which can even sometimes be fatal in these people. In animals, T. gondii infection can cause abortion or congenital anomalies. Three types of T. gondii I, II and III have been identified for the induction of virulence in mice as a laboratory animal model. Considering the high importance of molecular methods in terms of sensitivity, the target genes in this project are three major genes B1, GRA6 and REP-529. Among them B1 and REP-529 genes that are repeating genes throughout the genome, have 35 and 200 -300 copies in the genome of the parasite respectively.The reference specimens in this study are standard type I (RH strain), Type II (PRU strain) and Type III reference strains (VEG strain). Three types of T. gondii have grown on cell tissue culture separately and the cell concentrates were used for DNA extraction. The DNA was purified from proliferated parasites and it was used to carry out PCR experiments with the specific primers of the three mentioned genes. After PCR optimization, the PCR product undergoes PCR-RFLP method using AluI, XhoI and MseI restriction enzymes in terms of size of the product and digestion products in three types with all three primer pairs investigated and thus we will have a precise diagnostic model.

کلیدواژه ها:

Toxoplasma gondii ، Genotyping ، B1 ، Gra6 ، Rep529 ، PCR