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dc.contributor.author
Abras, Alba
dc.contributor.author
Ballart, Cristina
dc.contributor.author
Llovet, Teresa
dc.contributor.author
Roig, Carme
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Gutiérrez, Cristina
dc.contributor.author
Tebar, Silvia
dc.contributor.author
Berenguer, Pere
dc.contributor.author
Pinazo, María-Jesús
dc.contributor.author
Posada, Elizabeth
dc.contributor.author
Gascón, Joaquim
dc.contributor.author
Schijman, Alejandro Gabriel
dc.contributor.author
Gállego, Montserrat
dc.contributor.author
Muñoz, Carmen
dc.date.available
2019-07-18T19:47:53Z
dc.date.issued
2018-04
dc.identifier.citation
Abras, Alba; Ballart, Cristina; Llovet, Teresa; Roig, Carme; Gutiérrez, Cristina; et al.; Introducing automation to the molecular diagnosis of Trypanosoma cruzi infection: A comparative study of sample treatments, DNA extraction methods and real-time PCR assays; Public Library of Science; Plos One; 13; 4; 4-2018; 1-14
dc.identifier.issn
1932-6203
dc.identifier.uri
http://hdl.handle.net/11336/79837
dc.description.abstract
Background Polymerase chain reaction (PCR) has become a useful tool for the diagnosis of Trypanosoma cruzi infection. The development of automated DNA extraction methodologies and PCR systems is an important step toward the standardization of protocols in routine diagnosis. To date, there are only two commercially available Real-Time PCR assays for the routine laboratory detection of T. cruzi DNA in clinical samples: TCRUZIDNA.CE (Diagnostic Bioprobes Srl) and RealCycler CHAG (Progenie Molecular). Our aim was to evaluate the RealCycler CHAG assay taking into account the whole process. Methodology/Principal findings We assessed the usefulness of an automated DNA extraction system based on magnetic particles (EZ1 Virus Mini Kit v2.0, Qiagen) combined with a commercially available Real-Time PCR assay targeting satellite DNA (SatDNA) of T. cruzi (RealCycler CHAG), a methodology used for routine diagnosis in our hospital. It was compared with a well-known strategy combining a commercial DNA isolation kit based on silica columns (High Pure PCR Template Preparation Kit, Roche Diagnostics) with an in-house Real-Time PCR targeting SatDNA. The results of the two methodologies were in almost perfect agreement, indicating they can be used interchangeably. However, when variations in protocol factors were applied (sample treatment, extraction method and Real-Time PCR), the results were less convincing. A comprehensive fine-tuning of the whole procedure is the key to successful results. Guanidine EDTA-blood (GEB) samples are not suitable for DNA extraction based on magnetic particles due to inhibition, at least when samples are not processed immediately. Conclusions/Significance This is the first study to evaluate the RealCycler CHAG assay taking into account the overall process, including three variables (sample treatment, extraction method and Real-Time PCR). Our findings may contribute to the harmonization of protocols between laboratories and to a wider application of Real-Time PCR in molecular diagnostic laboratories associated with health centers.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Public Library of Science
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Real Time Pcr
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Automatic Dna Extraction
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Molecular Diagnosis
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Chagas Disease
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Bioquímica y Biología Molecular
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Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
dc.title
Introducing automation to the molecular diagnosis of Trypanosoma cruzi infection: A comparative study of sample treatments, DNA extraction methods and real-time PCR assays
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2019-07-17T13:13:20Z
dc.journal.volume
13
dc.journal.number
4
dc.journal.pagination
1-14
dc.journal.pais
Estados Unidos
dc.journal.ciudad
San Francisco
dc.description.fil
Fil: Abras, Alba. Universidad de Barcelona; España. Universidad de Girona; España. Instituto de Salud Global de Barcelona; España
dc.description.fil
Fil: Ballart, Cristina. Universidad de Barcelona; España. Instituto de Salud Global de Barcelona; España
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Fil: Llovet, Teresa. Universitat Autònoma de Barcelona; España. Hospital de la Santa Creu I Sant Pau; España
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Fil: Roig, Carme. Hospital de la Santa Creu I Sant Pau; España
dc.description.fil
Fil: Gutiérrez, Cristina. Hospital de la Santa Creu I Sant Pau; España
dc.description.fil
Fil: Tebar, Silvia. Universidad de Barcelona; España. Instituto de Salud Global de Barcelona; España
dc.description.fil
Fil: Berenguer, Pere. Hospital de la Santa Creu I Sant Pau; España
dc.description.fil
Fil: Pinazo, María-Jesús. Instituto de Salud Global de Barcelona; España
dc.description.fil
Fil: Posada, Elizabeth. Instituto de Salud Global de Barcelona; España
dc.description.fil
Fil: Gascón, Joaquim. Instituto de Salud Global de Barcelona; España
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: Gállego, Montserrat. Instituto de Salud Global de Barcelona; España. Universidad de Barcelona; España
dc.description.fil
Fil: Muñoz, Carmen. Hospital de la Santa Creu I Sant Pau; España. Universitat Autònoma de Barcelona; España
dc.journal.title
Plos One
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1371/journal.pone.0195738
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pubmed/29664973
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0195738
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