[Evaluation of TcH2AF-R and S35-S36 primers in PCR tests for the detection of Trypanosoma cruzi in mouse cardiac tissue].
BACKGROUND
Heart transplant is a therapeutic option in the treatment of chagasic cardiomyopathy. For early detection of Chagas reactivation cases, the use of PCR tests using endomyocardial biopsies has been proposed. Development of an animal model will be the first step in evaluating the applicability of this approach.


OBJECTIVE
PCR tests based on the TcH2AF-R and S35-S36 primers were evaluated for the detection of T. cruzi in heart tissue of mice experimentally infected with the parasite.


METHODS
Two groups of ICR mice of 15 and 10 individuals were infected by intraperitoneal injection with 0.3 ml of PBS containing 1 x 10(6) trypomastigotes of the MHOM/CO/2001/D.A. (T. cruzi I) strain or 1 x 10(4) trypomastigotes of MHOM/BR/00/Y (T. cruzi II) strain. Parasitemia and cardiac parasitic infection were determined at 30, 60 (acute model), 100 and 150 (chronic model) days by means of histopathological examination and by PCR, using the TcH2AF-R and S35-S36 primers.


RESULTS
The histopathological findings revealed alterations in the heart and the presence of intracellular amastigotes in acute and chronic models. In contrast to parasitemia levels and histopathological analyses, S35-S36 PCR detected infections in mice that were infected with either parasite strain. TcH2AF-R PCR detected T. cruzi I-infected mice earlier and more frequently than inspection for parasitemia or histopathological examination.


CONCLUSIONS
Applying PCR tests with both primers proved superior for Chagas disease confirmation over currently standard detection methods.
Barrera
Yuli Katherine
YK
Laboratorio de Parasitología Molecular, Departamento de Microbiología, Facultad de Ciencias, Pontificia Universidad Javeriana, Bogotá, D.C., Colombia.
Guevara
Johana María
JM
Pavía
Paula Ximena
PX
Montilla
Marleny
M
Nicholls
Rubén Santiago
RS
Parra
Edgar
E
Puerta
Concepción Judith
CJ
D000818:Animals; D002598:Chagas Cardiomyopathy / Q000469:parasitology / Q000473:pathology; D017931:DNA Primers / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics*; D006321:Heart / Q000469:parasitology*; D006801:Humans; D008297:Male; D051379:Mice; D009206:Myocardium / Q000166:cytology / Q000473:pathology; D016133:Polymerase Chain Reaction / Q000379:methods*; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000235:genetics*
D017931:DNA Primers; D016054:DNA, Protozoan; D015800:Protozoan Proteins