Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study
Rev. Soc. Bras. Med. Trop
;
35(5): 487-490, Sept.-Oct. 2002. ilus
Artículo
en Inglés
| LILACS
| ID: lil-328000
RESUMO
To evaluate the sensitivity of polymerase chain reaction (PCR) to reveal known number of trypomastigote in the blood of mice, three separate experiments were done. First To eight samples of 500mul of normal mice blood, one aliquot of 1, 2, 3, 4, 5, 10, and 50 trypomastigotes respectively, were added. Second and third 10 aliquots with 1 and 10 with 2 trypomastigotes were added to samples of 500mul of normal mice blood. Positive control 500mul of blood containing 100,000 trypomastigotes. For kDNA minicircles amplification by PCR the primersS35 and S36 were used. PCR revealed products of 330 b.p in the positive controls. When only one sample with the aliquots of 1 or 2 trypomastigotes was examined, results were negative; results were positive with aliquots of 3 to 50 trypomastigotes. In the 2nd and 3rd experiments, 9/10 aliquots with one parasite and 9/10 with 2 trypomastigotes were positive revealing a high sensitivity of this reaction. In conclusion, the presence of one single parasite in 500mul of blood, is enough for a positive PCR. This method could be used as a complement to the various parasitological cure tests in treated mice, when low volumes of blood are individually examined
Texto completo:
Disponible
Índice:
LILACS (Américas)
Asunto principal:
Trypanosoma cruzi
/
Reacción en Cadena de la Polimerasa
Tipo de estudio:
Estudio diagnóstico
Límite:
Animales
Idioma:
Inglés
Revista:
Rev. Soc. Bras. Med. Trop
Asunto de la revista:
Medicina Tropical
Año:
2002
Tipo del documento:
Artículo
País de afiliación:
Brasil
Institución/País de afiliación:
Fundaçäo Instituto Oswaldo Cruz/BR
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