Evaluation of real-time nucleic acid sequence-based amplification for detection of Chikungunya virus in clinical samples
Autor(es): Telles J-N, Le Roux K, Grivard P, Vernet G, Michault A
Resumo: The Chikungunya virus (CHIKV) is a member of the genus Alphavirus that is transmitted to humans by Aedes mosquitoes. In 2005 - 2006, the Indian Ocean isl- 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 - 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, - estimated to be 200 copies per NASBA reaction. The specificity of the assay was determined using blast analyses - non-cross-reactivity using an O'nyong-nyong virus culture - 250 CHIKV RT-PCR-negative plasma samples. A 100 % specificity was found - 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 - reliable method for the routine diagnosis of CHIKV in clinical samples.
Imprenta: Journal of Medical Microbiology, v. 58, Pt 9, p. 1168-1172, 2009
Identificador do objeto digital: 10.1099/jmm.0.010736-0
Descritores: Chikungunya virus - Molecular structure ; Chikungunya virus - Pathogenesis ; Chikungunya virus - RNA ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya virus - PCR detection ; Chikungunya virus - Real Time PCR ; Chikungunya virus - RT-PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Epidemiology ; Chikungunya virus - Public health
Data de publicação: 2009
