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J Med Microbiol 58 (2009), 1168-1172; DOI: 10.1099/jmm.0.010736-0
© 2009 Society for General Microbiology
ISSN 0022-2615

Evaluation of real-time nucleic acid sequence-based amplification for detection of Chikungunya virus in clinical samples

J.-N. Telles1,2, K. Le Roux3, P. Grivard3, G. Vernet1,2 and A. Michault3

1 bioMérieux, 69280 Marcy L'Etoile, France

2 Fondation Mérieux, Département des Pathogènes Emergents, 69007 Lyon, France

3 Laboratoire de Bactériologie-Parasitologie-Virologie et Hygiène, Groupe Hospitalier Sud Réunion, Centre Hospitalier Régional, 97448 Saint Pierre, La Réunion, France

Correspondence
A. Michault
a.michault{at}chr-reunion.fr

Received February 25, 2009
Accepted May 31, 2009

The Chikungunya virus (CHIKV) is a member of the genus Alphavirus that is transmitted to humans by Aedes mosquitoes. In 2005 and 2006, the Indian Ocean island of La Réunion was hit with an unprecedented CHIKV fever outbreak that infected 300 000 people. In the present study, we describe the evaluation of real-time nucleic acid sequence-based amplification (RT-NASBA) for the detection of CHIKV in clinical samples. A co-extracted and co-amplified chimerical CHIKV RNA sequence was used as an internal control to eliminate false-negative results. The detection threshold of the assay was determined from quantified CHIKV-positive plasma, and estimated to be 200 copies per NASBA reaction. The specificity of the assay was determined using BLAST analyses and non-cross-reactivity using an O'nyong-nyong virus culture and 250 CHIKV RT-PCR-negative plasma samples. A 100 % specificity was found and no invalid result was obtained, showing the good quality of the nucleic acid extraction. The assay was then evaluated using 252 CHIKV-positive RT-PCR plasma samples. The samples were all tested positive, including those with low viral load. This evaluation showed that the RT-NASBA is a rapid (5 h from sample nucleic acid extraction to detection), sensitive, specific and reliable method for the routine diagnosis of CHIKV in clinical samples.






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