Autor(es): Saha Kallol, Firdaus Rushna, Chakrabarti Sekhar, Sadhukhan Provash Chandra

Resumo: Chikungunya - dengue, two arboviral infections are common in South-East Asia - their early clinical manifestations are very similar hence it is important to discriminate between them as early as possible for better clinical management. The aim of this study was to design a rapid, sensitive - specific method for the differential diagnosis of these two viruses simultaneously. A rapid one-tube duplex RT-PCR assay was developed that requires 110 min including RNA extraction, RT-PCR - agarose gel electrophoresis by using a novel Taq polymerase with high processivity. This one-tube duplex RT-PCR system with primers designed from the conserved regions of the genome allowed discrimination between the two viral groups. Bioinformatics analysis of the DNA sequences from PCR amplified products confirmed that this method was very specific - accurate. The time required for this duplex RT-PCR was comparable to the st-ard IgM capture ELISA method. This novel approach would help to diagnose specifically - accurately these two closely related arboviruses - enable early detection from blood. This method could be applied in resource limited settings, for surveillance in endemic regions or for routine epidemiological screening.

Palavras-Chave: Arboviruses; Chikungunya; Dengue; Duplex RT-PCR; Rapid Nucleic Acid tests

Imprenta: Journal of Virological Methods, v. 193, n. 2, p. 521-514, 2013

Identificador do objeto digital: 10.1016/j.jviromet.2013.07.029

Descritores: Chikungunya virus - Arbovirus ; Chikungunya virus - DNA ; Chikungunya virus - Flaviviridae ; Chikungunya virus - Genome ; Chikungunya virus - Pathogenesis ; Chikungunya virus - RNA ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya virus - PCR detection ; 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: 2013