This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Garcia, S.
Right arrow Articles by Garin, D.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Garcia, S.
Right arrow Articles by Garin, D.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, December 2001, p. 4456-4461, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4456-4461.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Quantitative Real-Time PCR Detection of Rift Valley Fever Virus and Its Application to Evaluation of Antiviral Compounds

Stephan Garcia,1 Jean Marc Crance,1 Agnes Billecocq,2 Andre Peinnequin,1 Alain Jouan,1 Michele Bouloy,2 and Daniel Garin1,*

Unité de Virologie, Centre de Recherches du Service de Santé des Armées (CRSSA) Emile Pardé, Grenoble,1 and Groupe des Bunyaviridés, Institut Pasteur, Paris,2 France

Received 6 August 2001/Returned for modification 12 September 2001/Accepted 27 September 2001

The Rift Valley fever virus (RVFV), a member of the genus Phlebovirus (family Bunyaviridae) is an enveloped negative-strand RNA virus with a tripartite genome. Until 2000, RVFV circulation was limited to the African continent, but the recent deadly outbreak in the Arabian Peninsula dramatically illustrated the need for rapid diagnostic methods, effective treatments, and prophylaxis. A method for quantifying the small RNA segment by a real-time detection reverse transcription (RT)-PCR using TaqMan technology and targeting the nonstructural protein-coding region was developed, and primers and a probe were designed. After optimization of the amplification reaction and establishment of a calibration curve with synthetic RNA transcribed in vitro from a plasmid containing the gene of interest, real-time RT-PCR was assessed with samples consisting of RVFV from infected Vero cells. The method was found to be specific for RVFV, and it was successfully applied to the detection of the RVFV genome in animal sera infected with RVFV as well as to the assessment of the efficiency of various drugs (ribavirin, alpha interferon, 6-azauridine, and glycyrrhizin) for antiviral activity. Altogether, the results indicated a strong correlation between the infectious virus titer and the amount of viral genome assayed by real time RT-PCR. This novel method could be of great interest for the rapid diagnosis and screening of new antiviral compounds, as it is sensitive and time saving and does not require manipulation of infectious material.

* Corresponding author. Mailing address: Unité de Virologie, CRSSA, 24 avenue des Maquis du Grésivaudan, BP 87-38702 La Tronche cedex, France. Phone: 33-476-63-68-44. Fax: 33-476-63-69-17. E-mail: Daniel.Garin{at}wanadoo.fr.

Journal of Clinical Microbiology, December 2001, p. 4456-4461, Vol. 39, No. 12
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.12.4456-4461.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Le Roux, C. A., Kubo, T., Grobbelaar, A. A., van Vuren, P. J., Weyer, J., Nel, L. H., Swanepoel, R., Morita, K., Paweska, J. T. (2009). Development and Evaluation of a Real-Time Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Rift Valley Fever Virus in Clinical Specimens. J. Clin. Microbiol. 47: 645-651 [Abstract] [Full Text]  
  • Peyrefitte, C. N., Boubis, L., Coudrier, D., Bouloy, M., Grandadam, M., Tolou, H. J., Plumet, S. (2008). Real-Time Reverse-Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of Rift Valley Fever Virus. J. Clin. Microbiol. 46: 3653-3659 [Abstract] [Full Text]  
  • Bird, B. H., Bawiec, D. A., Ksiazek, T. G., Shoemaker, T. R., Nichol, S. T. (2007). Highly Sensitive and Broadly Reactive Quantitative Reverse Transcription-PCR Assay for High-Throughput Detection of Rift Valley Fever Virus. J. Clin. Microbiol. 45: 3506-3513 [Abstract] [Full Text]  
  • Scaramozzino, N., Ferrier-Rembert, A., Favier, A.-l., Rothlisberger, C., Richard, S., Crance, J.-M., Meyer, H., Garin, D. (2007). Real-Time PCR to Identify Variola Virus or Other Human Pathogenic Orthopox Viruses. Clin. Chem. 53: 606-613 [Abstract] [Full Text]  
  • KEMPF, B. J., BLAIR, C. D., BEATY, B. J. (2006). QUANTITATIVE ANALYSIS OF LA CROSSE VIRUS TRANSCRIPTION AND REPLICATION IN CELL CULTURES AND MOSQUITOES. Am J Trop Med Hyg 74: 224-232 [Abstract] [Full Text]  
  • Oozeer, R., Furet, J. P., Goupil-Feuillerat, N., Anba, J., Mengaud, J., Corthier, G. (2005). Differential Activities of Four Lactobacillus casei Promoters during Bacterial Transit through the Gastrointestinal Tracts of Human-Microbiota-Associated Mice. Appl. Environ. Microbiol. 71: 1356-1363 [Abstract] [Full Text]  
  • Hughes, G. J., Smith, J. S., Hanlon, C. A., Rupprecht, C. E. (2004). Evaluation of a TaqMan PCR Assay To Detect Rabies Virus RNA: Influence of Sequence Variation and Application to Quantification of Viral Loads. J. Clin. Microbiol. 42: 299-306 [Abstract] [Full Text]  
  • Drosten, C., Gottig, S., Schilling, S., Asper, M., Panning, M., Schmitz, H., Gunther, S. (2002). Rapid Detection and Quantification of RNA of Ebola and Marburg Viruses, Lassa Virus, Crimean-Congo Hemorrhagic Fever Virus, Rift Valley Fever Virus, Dengue Virus, and Yellow Fever Virus by Real-Time Reverse Transcription-PCR. J. Clin. Microbiol. 40: 2323-2330 [Abstract] [Full Text]  
  • Sall, A. A., Macondo, E. A., Sene, O. K., Diagne, M., Sylla, R., Mondo, M., Girault, L., Marrama, L., Spiegel, A., Diallo, M., Bouloy, M., Mathiot, C. (2002). Use of Reverse Transcriptase PCR in Early Diagnosis of Rift Valley Fever. CVI 9: 713-715 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»