From each participant, we collected 20 mL of EDTA-anti-coagulated blood that were immediately mixed with an equal volume of 6 M guanidine hydrochloride -0.2M EDTA solution. The target-capture real-time PCR assay used in this study was developed based on the PCR method described by Virreira et al.,7 targeting the kinetoplast T. cruzi mitochondrial minicircle DNA. The DNA extraction was improved with a target-capture step using magnetic beads coated with a T. cruzi-specific 20-mer capture oligonucleotide.

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