Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study
Autor(a) principal: | |
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Data de Publicação: | 2002 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista da Sociedade Brasileira de Medicina Tropical |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822002000500011 |
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 primers:S35 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. |
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Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro studyTrypanosoma cruziPolymerase chain reactionPCR sensitivity in micePCR as cure test in miceTo 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 primers:S35 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.Sociedade Brasileira de Medicina Tropical - SBMT2002-10-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822002000500011Revista da Sociedade Brasileira de Medicina Tropical v.35 n.5 2002reponame:Revista da Sociedade Brasileira de Medicina Tropicalinstname:Sociedade Brasileira de Medicina Tropical (SBMT)instacron:SBMT10.1590/S0037-86822002000500011info:eu-repo/semantics/openAccessCampos,Rozália F.Magalhães,Juracy B.Reis,Eliana A.G.Reis,Mitermayer G.Andrade,Sonia G.eng2002-11-27T00:00:00Zoai:scielo:S0037-86822002000500011Revistahttps://www.sbmt.org.br/portal/revista/ONGhttps://old.scielo.br/oai/scielo-oai.php||dalmo@rsbmt.uftm.edu.br|| rsbmt@rsbmt.uftm.edu.br1678-98490037-8682opendoar:2002-11-27T00:00Revista da Sociedade Brasileira de Medicina Tropical - Sociedade Brasileira de Medicina Tropical (SBMT)false |
dc.title.none.fl_str_mv |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study |
title |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study |
spellingShingle |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study Campos,Rozália F. Trypanosoma cruzi Polymerase chain reaction PCR sensitivity in mice PCR as cure test in mice |
title_short |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study |
title_full |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study |
title_fullStr |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study |
title_full_unstemmed |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study |
title_sort |
Sensitivity of polymerase chain reaction for detection of known aliquots of Trypanosoma cruzi in the blood of mice: an in vitro study |
author |
Campos,Rozália F. |
author_facet |
Campos,Rozália F. Magalhães,Juracy B. Reis,Eliana A.G. Reis,Mitermayer G. Andrade,Sonia G. |
author_role |
author |
author2 |
Magalhães,Juracy B. Reis,Eliana A.G. Reis,Mitermayer G. Andrade,Sonia G. |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Campos,Rozália F. Magalhães,Juracy B. Reis,Eliana A.G. Reis,Mitermayer G. Andrade,Sonia G. |
dc.subject.por.fl_str_mv |
Trypanosoma cruzi Polymerase chain reaction PCR sensitivity in mice PCR as cure test in mice |
topic |
Trypanosoma cruzi Polymerase chain reaction PCR sensitivity in mice PCR as cure test in mice |
description |
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 primers:S35 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. |
publishDate |
2002 |
dc.date.none.fl_str_mv |
2002-10-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822002000500011 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0037-86822002000500011 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0037-86822002000500011 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Medicina Tropical - SBMT |
publisher.none.fl_str_mv |
Sociedade Brasileira de Medicina Tropical - SBMT |
dc.source.none.fl_str_mv |
Revista da Sociedade Brasileira de Medicina Tropical v.35 n.5 2002 reponame:Revista da Sociedade Brasileira de Medicina Tropical instname:Sociedade Brasileira de Medicina Tropical (SBMT) instacron:SBMT |
instname_str |
Sociedade Brasileira de Medicina Tropical (SBMT) |
instacron_str |
SBMT |
institution |
SBMT |
reponame_str |
Revista da Sociedade Brasileira de Medicina Tropical |
collection |
Revista da Sociedade Brasileira de Medicina Tropical |
repository.name.fl_str_mv |
Revista da Sociedade Brasileira de Medicina Tropical - Sociedade Brasileira de Medicina Tropical (SBMT) |
repository.mail.fl_str_mv |
||dalmo@rsbmt.uftm.edu.br|| rsbmt@rsbmt.uftm.edu.br |
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1752122151907885056 |