In vitro transcribed RNA molecules for the diagnosis of pandemic 2009 influenza A(H1N1) virus by real-time RT-PCR

Mario Bermúdez de León, Katia Peñuelas-Urquides, Miguel E. Aguado-Barrera, María José Currás-Tuala, Brenda L. Escobedo-Guajardo, Rosa Nelly González-Ríos, Viviana L. Mata-Tijerina, Ofelia E. Vázquez-Monsiváis

Resultado de la investigaciónrevisión exhaustiva

2 Citas (Scopus)


The 2009 influenza A(H1N1) outbreak allowed the implementation of new epidemiologic surveillance tools in several countries around the world. A new molecular protocol with appropriate sensitivity and specificity using real-time RT-PCR was developed by the Centers for Disease Control and Prevention (CDC) to identify the pandemic 2009 influenza A (H1N1) virus in human specimens. In the CDC protocol, positive controls are available only upon request and they are taken from cell cultures infected with 2009 influenza A(H1N1) virus, representing a handling risk for laboratory technicians. The poor availability of positive control materials in diagnostic laboratories may limit the public health response. The aim of the work presented in this paper was to develop positive controls for the diagnostic testing of influenza A(H1N1) virus that could be used in the CDC real-time RT-PCR protocol. A series of plasmid constructs bearing partial sequences of the viral genes were created and each construct was used as a template for in vitro transcription. RNA molecules were obtained successfully at high yield, i.e., 2×10 7 assays per microliter. Thus, the inclusion of these molecules in the influenza panel as positive controls is proposed. The in vitro transcribed RNA could also be used as quality standards in the design of international proficiency studies.

Idioma originalEnglish
Páginas (desde-hasta)487-491
Número de páginas5
PublicaciónJournal of Virological Methods
EstadoPublished - 1 nov. 2013
Publicado de forma externa

All Science Journal Classification (ASJC) codes

  • Virología


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