Evaluation of the inhibitory effects of drugs on the growth of Babesia Parasites using real-time PCR method

Abstract

In order to evaluate the inhibitory effects of drug on the growth of babesia parasite, relative quantification real-time PCR method was developed in this study. The 18S rRNA gene was used as target gene for the 2-{Delta}{Delta}Ct method analysis. Meanwhile, Chicken RNA was added into the parasitized blood for total RNA extraction. The {beta}-actin gene of chicken was selected as internal control gene for the 2-{Delta}{Delta}Ct method analysis. Parasitized blood 100 L, 50 L, 25L, 12.5 L, 6.25 L was prepared for B. gibsoni relative quantification. Regression analysis results revealed that significant linear relationships between the relative quantification value and parasitemia. The 18S rRNA gene expression was significantly decreased after the treatment of Diminazene aceturate and Artesunate in vitro drug sensitivity test. It suggested that this relative quantification real-time PCR method can be used in evaluating the effects of inhibitory of drug.