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
Artigo
em Inglês
| 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:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Trypanosoma cruzi
/
Reação em Cadeia da Polimerase
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
Idioma:
Inglês
Revista:
Rev. Soc. Bras. Med. Trop
Assunto da revista:
Medicina Tropical
Ano de publicação:
2002
Tipo de documento:
Artigo
País de afiliação:
Brasil
Instituição/País de afiliação:
Fundaçäo Instituto Oswaldo Cruz/BR
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