Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples

Aims: Molecular biology techniques based on the detection of genomic sequences by reverse transcription combined with polymerase chain reaction (PCR) have enabled the detection of different RNA viruses in serum or plasma samples. Since the dengue epidemic outbreak declared in Argentina in 2009, nume...

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Autores principales: Galván, Cristian Ariel, Elbarcha, Osvaldo César, Fernández, Eduardo J., Beltramo, Dante Miguel, Soria, Néstor Walter
Formato: Artículo PeerReviewed
Lenguaje:Español
Publicado: 2011
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QD Química
Acceso en línea:http://pa.bibdigital.ucc.edu.ar/5016/1/A_Galv%C3%A1n.pdf
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Producción Académica Universidad Católica de Córdoba (UCCor) de Universidad Católica de Córdoba
id I38-R144-5016
record_format dspace
spelling I38-R144-50162025-11-11T17:28:08Z http://pa.bibdigital.ucc.edu.ar/5016/ Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples Galván, Cristian Ariel Elbarcha, Osvaldo César Fernández, Eduardo J. Beltramo, Dante Miguel Soria, Néstor Walter QD Química Aims: Molecular biology techniques based on the detection of genomic sequences by reverse transcription combined with polymerase chain reaction (PCR) have enabled the detection of different RNA viruses in serum or plasma samples. Since the dengue epidemic outbreak declared in Argentina in 2009, numerous patients' samples were analyzed for the acute phase of infection. One of the main methodological drawbacks is the lack of internal control to measure the effectiveness of the viral extraction and reverse transcription process. In this article, we propose to standardize a molecular method to detect beta actin (β-Act) and glucose 6 phosphate dehydrogenase (G6PDH) complementary DNAs (cDNAs) present in patient's plasma/serum, as a control process. Results: RNA extraction, reverse transcription, and PCRs for human G6PDH, β-Act, and the dengue virus genome were performed. cDNA fragments for β-Act and G6PDH were amplified for all samples, regardless of the presence or absence of viral RNA. Conclusions: Amplification of β-Act and G6PDH cDNAs can be used as a control for the extraction and reverse transcription processes during dengue virus detection. This could also be a useful method for controlling the above steps when infections caused by other RNA viruses are studied, even if another methodology is employed, such as real-time PCR. 2011-12-31 Artículo PeerReviewed application/pdf spa http://pa.bibdigital.ucc.edu.ar/5016/1/A_Galv%C3%A1n.pdf Galván, Cristian Ariel ORCID: https://orcid.org/0000-0002-1885-3664 <https://orcid.org/0000-0002-1885-3664>, Elbarcha, Osvaldo César, Fernández, Eduardo J., Beltramo, Dante Miguel ORCID: https://orcid.org/0000-0002-8198-3525 <https://orcid.org/0000-0002-8198-3525> and Soria, Néstor Walter ORCID: https://orcid.org/0000-0002-8232-1822 <https://orcid.org/0000-0002-8232-1822> (2011) Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples. Genetic Testing and Molecular Biomarkers, 15 (12). pp. 913-915. ISSN 19450257 info:eu-repo/semantics/altIdentifier/doi/10.1089/gtmb.2011.0091
institution Universidad Católica de Córdoba
institution_str I-38
repository_str R-144
collection Producción Académica Universidad Católica de Córdoba (UCCor)
language Español
orig_language_str_mv spa
topic QD Química
spellingShingle QD Química
Galván, Cristian Ariel
Elbarcha, Osvaldo César
Fernández, Eduardo J.
Beltramo, Dante Miguel
Soria, Néstor Walter
Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples
topic_facet QD Química
description Aims: Molecular biology techniques based on the detection of genomic sequences by reverse transcription combined with polymerase chain reaction (PCR) have enabled the detection of different RNA viruses in serum or plasma samples. Since the dengue epidemic outbreak declared in Argentina in 2009, numerous patients' samples were analyzed for the acute phase of infection. One of the main methodological drawbacks is the lack of internal control to measure the effectiveness of the viral extraction and reverse transcription process. In this article, we propose to standardize a molecular method to detect beta actin (β-Act) and glucose 6 phosphate dehydrogenase (G6PDH) complementary DNAs (cDNAs) present in patient's plasma/serum, as a control process. Results: RNA extraction, reverse transcription, and PCRs for human G6PDH, β-Act, and the dengue virus genome were performed. cDNA fragments for β-Act and G6PDH were amplified for all samples, regardless of the presence or absence of viral RNA. Conclusions: Amplification of β-Act and G6PDH cDNAs can be used as a control for the extraction and reverse transcription processes during dengue virus detection. This could also be a useful method for controlling the above steps when infections caused by other RNA viruses are studied, even if another methodology is employed, such as real-time PCR.
format Artículo
PeerReviewed
author Galván, Cristian Ariel
Elbarcha, Osvaldo César
Fernández, Eduardo J.
Beltramo, Dante Miguel
Soria, Néstor Walter
author_facet Galván, Cristian Ariel
Elbarcha, Osvaldo César
Fernández, Eduardo J.
Beltramo, Dante Miguel
Soria, Néstor Walter
author_sort Galván, Cristian Ariel
title Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples
title_short Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples
title_full Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples
title_fullStr Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples
title_full_unstemmed Development of a method to control the RNA extraction and reverse transcription steps for the detection of dengue virus present in human blood samples
title_sort development of a method to control the rna extraction and reverse transcription steps for the detection of dengue virus present in human blood samples
publishDate 2011
url http://pa.bibdigital.ucc.edu.ar/5016/1/A_Galv%C3%A1n.pdf
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