Simultaneous detection and quantitation of Chikungunya, dengue and West Nile viruses by multiplex RT-PCR assays and Dengue virus typing using high resolution melting
Autor(es): Naze F, Le Roux K, Schuffenecker I, Zeller H, Staikowsky F, Grivard P, Michault A, Laurent P
Resumo: Chikungunya (CHIKV), Dengue (DENV) - West Nile (WNV) viruses are arthropod-borne viruses that are able to emerge or re-emerge in many regions due to climatic changes - increase in travel. Since these viruses produce similar clinical signs it is important for physicians - epidemiologists to differentiate them rapidly. A molecular method was developed for their detection - quantitation in plasma samples - a DENV typing technique were developed. The method consisted in performing two multiplex real-time one-step RT-PCR assays, to detect - quantify the three viruses. Both assays were conducted in a single run, from a single RNA extract containing a unique coextracted - coamplified composite internal control. The quantitation results were close to the best detection thresholds obtained with simplex RT-PCR techniques. The differentiation of DENV types was performed using a High Resolution Melting technique. The assays enable the early diagnosis of the three arboviruses during viremia, including cases of coinfection. The method is rapid, specific - highly sensitive with a potential for clinical diagnosis - epidemiological surveillance. A DENV positive sample can be typed conveniently using the High Resolution Melting technique using the same apparatus.
Palavras-Chave: Multiplex real-time RT-PCR; Arboviruses; Absolute quantitation; Internal control; Dengue virus typing; High Resolution Melting
Imprenta: Journal of Virological Methods, v. 162, n. 1-2, p. 1-7, 2009
Identificador do objeto digital: 10.1016/j.jviromet.2009.03.006
Descritores: Chikungunya virus - Arbovirus ; Chikungunya virus - Flaviviridae ; 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 - Dengue ; Chikungunya virus - Public health
Data de publicação: 2009
