PCR Amplification Of Trypanosoma Cruzi - Specific DNA from Raccoon Blood Samples

Colin Moss, Western Kentucky University

Abstract

The parasitic protist Trypanosoma cruzi is the causative agent of Chagas Disease. Chagas Disease causes greater than 15,000 deaths each year, and nearly 28 million people are believed to be at risk of infection in Central and South America. This parasite has been described in many mammalian host species and has also been described in the United States. The purpose of this study was to attempt to use PCR to amplify T. cruzi-specific DNA directly from blood samples obtained from raccoons (Procyon lotor) trapped in Warren and Barren Counties of Kentucky in 2007 and 2008. DNA was successfully isolated from 487raccoon blood samples using a Qiagen QIAmp DNA Blood Mini Kit. Each DNA sample was then subjected to PCR amplification using the T. cruzi specific primer pair known as TCZ1 and TCZ2. It was determined that T. cruzi DNA was present in 47% of raccoon blood samples with a 57% prevalence in Warren county and a 32% prevalence in Barren county. Groce (2008) estimated a similar overall prevalence of 38% in these same raccoons based upon the results of hemocultures established at the time of sample collection.