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Polymerase Chain Reaction Detection of Trypanosoma cruzi in Macaca fascicularis Using Archived Tissues

This study describes conventional and real-time polymerase chain reaction (PCR) methods developed to detect and quantify Trypanosoma cruzi DNA in cynomolgus monkeys (Macaca fascicularis) using formalin-fixed paraffin-embedded blocks archived for periods of up to 6 years. The highest concentration of T. cruzi DNA was found in the myocardium, urinary bladder, stomach, lymph node, adrenal gland, and colon. The concentration of T. cruzi DNA detected in cardiac tissues was 10–100-fold greater than found elsewhere; the mean concentrations of T. cruzi DNA in non-cardiac tissues were otherwise comparable. Trypanosoma cruzi DNA was amplified from cerebrum but not cerebellum or kidney. Successful use of DNA from formalin-fixed, paraffin-embedded blocks is important because most pathology laboratories routinely archive wax blocks. This archived resource can be used for further studies on the prevalence of this disease.

Received January 12, 2009. Accepted for publication May 7, 2009.

Acknowledgments: We gratefully acknowledge the technical assistance of Mary Jo Aivaliotis, Debbie Christian, Allen Ford, Sam Galindo, Janice MacRossin, Michael Torres, and Jane VandeBerg in collecting, processing, and assaying samples; Marie Silva, Antonio Perez for histopathology support; and Marie Tehas for photographic support. Mary Sparks provided expert assistance with data management and animal records research. We are also indebted to Tim Anderson, Nico Gouin, and Susan Mahaney for many helpful discussions.

Financial support: This research was supported in part by National Institutes of Health grants R01 RR016347 (to JTW) and P51 RR013986, and in part from facilities constructed with support from Research Facilities Improvement Program grants C06 RR015456 and C06 RR014578 from the National Center for Research Resources, National Institutes of Health.

^* Address correspondence to James N. Mubiru, Southwest Foundation for Biomedical Research, PO Box 760549, San Antonio, TX 78245-0549. E-mail: jmubiru{at}sfbr.org

Authors’ addresses: Jeff T. Williams, James N. Mubiru, Rohina C. Rubicz, John L. VandeBerg, Edward J. Dick Jr, and Gene B. Hubbard, Southwest Foundation for Biomedical Research, PO Box 760549, San Antonio, TX 78245-0549, Tel: 210-258-9400, Fax: 210-670-3323. Natalia E. Schlabritz-Loutsevitch, Department of Obstetrics and Gynecology, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78229.

Reprint requests: James N. Mubiru, Southwest Foundation for Biomedical Research, PO Box 760549, San Antonio, TX 78245-0549, E-mail: jmubiru{at}sfbr.org.