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dc.contributor.author
Qvarnstrom, Yvonne
dc.contributor.author
Schijman, Alejandro Gabriel
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dc.contributor.author
Veron, Vincent
dc.contributor.author
Aznar, Christine
dc.contributor.author
Steurer, Francis
dc.contributor.author
da Silva, Alexandre J.
dc.date.available
2020-05-13T16:34:08Z
dc.date.issued
2012-07
dc.identifier.citation
Qvarnstrom, Yvonne; Schijman, Alejandro Gabriel; Veron, Vincent; Aznar, Christine; Steurer, Francis; et al.; Sensitive and Specific Detection of Trypanosoma cruzi DNA in Clinical Specimens Using a Multi-Target Real-Time PCR Approach; Public Library of Science; Neglected Tropical Diseases; 6; 7; 7-2012; 1-8
dc.identifier.issn
1935-2735
dc.identifier.uri
http://hdl.handle.net/11336/105043
dc.description.abstract
Background: The laboratory diagnosis of Chagas disease is challenging because the usefulness of different diagnostic tests will depend on the stage of the disease. Serology is the preferred method for patients in the chronic phase, whereas PCR can be successfully used to diagnose acute and congenital cases. Here we present data using a combination of three TaqMan PCR assays to detect T. cruzi DNA in clinical specimens. Methods/Principal Findings: Included in the analysis were DNA extracted from 320 EDTA blood specimens, 18 heart tissue specimens, 6 umbilical cord blood specimens, 2 skin tissue specimens and 3 CSF specimens. For the blood specimens both whole blood and buffy coat fraction were analyzed. The specimens were from patients living in the USA, with suspected exposure to T. cruzi through organ transplantation, contact with triatomine bugs or laboratory accidents, and from immunosuppressed patients with suspected Chagas disease reactivation. Real-time PCR was successfully used to diagnose acute and Chagas disease reactivation in 20 patients, including one case of organ-transmitted infection and one congenital case. Analysis of buffy coat fractions of EDTA blood led to faster diagnosis in six of these patients compared to whole blood analysis. The three real-time PCR assays produced identical results for 94% of the specimens. The major reason for discrepant results was variable sensitivity among the assays, but two of the real-time PCR assays also produced four false positive results. Conclusions/Significance: These data strongly indicate that at least two PCR assays with different performances should be combined to increase the accuracy. This evaluation also highlights the benefit of extracting DNA from the blood specimen’s buffy coat to increase the sensitivity of PCR analysis.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Public Library of Science
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
POLYMERASE CHAIN REACTION
dc.subject
CHAGAS DISEASE
dc.subject
MOLECULAR DIAGNOSIS
dc.subject.classification
Bioquímica y Biología Molecular
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dc.subject.classification
Ciencias Biológicas
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dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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dc.title
Sensitive and Specific Detection of Trypanosoma cruzi DNA in Clinical Specimens Using a Multi-Target Real-Time PCR Approach
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2020-04-23T19:17:59Z
dc.journal.volume
6
dc.journal.number
7
dc.journal.pagination
1-8
dc.journal.pais
Estados Unidos
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dc.journal.ciudad
San Francisco
dc.description.fil
Fil: Qvarnstrom, Yvonne. Centers for Disease Control and Prevention; Estados Unidos
dc.description.fil
Fil: Schijman, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular "Dr. Héctor N. Torres"; Argentina
dc.description.fil
Fil: Veron, Vincent. Universitaire CH Andrée Rosemon; Francia
dc.description.fil
Fil: Aznar, Christine. Universitaire CH Andrée Rosemon; Francia
dc.description.fil
Fil: Steurer, Francis. Centers for Disease Control and Prevention; Estados Unidos
dc.description.fil
Fil: da Silva, Alexandre J.. Centers for Disease Control and Prevention; Estados Unidos
dc.journal.title
Neglected Tropical Diseases
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.plosntds.org/article/info%3Adoi%2F10.1371%2Fjournal.pntd.0001689
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/https://doi.org/10.1371/journal.pntd.0001689
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