Twenty-nine road-killed opossums (Didelphis virginiana) collected from Baldwin County, central Georgia, were tested for the presence of Trypanosoma cruzi with polymerase chain reaction (PCR) analysis. Utilizing a cell culture containing T. cruzi epimastigotes, a positive control was established by injecting parasites into opossum heart tissue. DNA was extracted from both the control and sample heart tissues using a proteinase K protocol. PCR analysis of the isolated DNA with T. cruzi-specific primers indicated the presence of parasite DNA in at least 10.3% of the opossums in the Baldwin County sample. This study demonstrates that PCR of extracted DNA from road-killed specimens is an effective method for detecting the presence of T. cruzi in opossums.

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