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Journal of Clinical Microbiology, November 2008, p. 3653-3659, Vol. 46, No. 11
0095-1137/08/$08.00+0     doi:10.1128/JCM.01188-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Real-Time Reverse-Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of Rift Valley Fever Virus{triangledown}

Christophe N. Peyrefitte,1 Laetitia Boubis,1 Daniel Coudrier,2 Michèle Bouloy,2 Marc Grandadam,2 Hugues J. Tolou,1 and Sébastien Plumet1*

Unité de virologie tropicale, Institut de Médecine tropicale du Service de Santé des Armées, BP 46, 13 998 Marseille armées, France,1 Unité de génétique moléculaire des Bunyavirus, Institut Pasteur, 25-28 rue du Dr Roux, 75724 Paris, Cedex 15, France2

Received 23 June 2008/ Returned for modification 29 July 2008/ Accepted 8 September 2008

The development and validation of a one-step, single-tube, real-time accelerated reverse-transcription loop-mediated isothermal amplification (RT-LAMP) for the detection of the L RNA segment of Rift Valley fever virus (RVFV) are described. The assay was performed at a constant temperature (63°C), with a real-time follow-up using a LightCycler and a double-stranded-DNA-intercalating fluorochrome. The assay is highly sensitive and comparable to real-time RT-PCR, with a detection limit of ~10 RNA copies per assay. However, the RT-LAMP assay is much faster than traditional RT-PCR and generates results in <30 min for most diluted samples. The specificity of the primers was established using other, related arboviruses as well as virus-containing and virus-free sera. The RT-LAMP assay reported here is thus a valuable tool for the rapid detection of RVFV in field diagnostic laboratories.

* Corresponding author. Mailing address: Unité de virologie tropicale, Institut de Médecine tropicale du Service de Santé des Armées, BP 46, 13 998 Marseille armées, France. Phone: 33-491-15-01-18. Fax: 33-491-15-01-72. E-mail: sebastien.plumet{at}m4x.org

{triangledown} Published ahead of print on 17 September 2008.

Journal of Clinical Microbiology, November 2008, p. 3653-3659, Vol. 46, No. 11
0095-1137/08/$08.00+0     doi:10.1128/JCM.01188-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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