Improvement of ITS1-PCR system by using FTA~R cards and Ampdirect~R Plus buffer for detection of Trypanosoma evansi infection

摘要

Trypanosoma evansi is one of the most widely distributed hemoparasites in Asia, and its infection of ruminants is listed up among 13 priority animal diseases in Indonesia. For epidemiological studies on T. evansi infection, internal transcribed spacer (ITS) 1 of the ribosomal DNA has been a target of PCR amplification, and a convenient PCR system with FTA~R cards (Whatman) has been reported. However, this system requires a time-consuming step of washing DNA samples spotted on FTA punches, and actual sensitivity has not yet been determined. This study describes the improvement of ITSI-PCR by using a combination of FTA~R cards and Ampdirect~R Plus buffer (Shimadzu), special PCR buffer capable of inactivating inhibitory factors in biological samples. Parasite stock was inoculated into rats, and blood samples were collected from the rats developing high parasitemia as well as non-infected rats.