Clinical evaluation of a single-reaction real-time RT-PCR for pan-dengue and chikungunya virus detection
Autor(es): Waggoner Jesse J, Ballesteros Gabriela, Gresh Lionel, Mohamed-Hadley Alisha, Tellez Yolanda, Sahoo Malaya K, Abeynayake Janaki, Balmaseda Angel, Harris Eva, Pinsky Benjamin A
Resumo: Dengue virus (DENV) - chikungunya virus (CHIKV) now co-circulate throughout tropical regions of the world, with billions of people living at risk of infection. The differentiation of these infections is important for epidemiologic surveillance as well as clinical care, though widely-used molecular diagnostics for DENV - CHIKV require the performance of two to four separate PCR reactions for detection. In the current study, we sought to develop - evaluate a single-reaction, multiplex real-time RT-PCR (rRT-PCR) for the detection - differentiation of DENV - CHIKV (the pan-DENV-CHIKV rRT-PCR). From an alignment of all available CHIKV complete genome sequences in GenBank, a new CHIKV rRT-PCR was designed for use in multiplex with a previously described assay for pan-DENV detection. Analytical evaluation was performed in accordance with published recommendations, - the pan-DENV-CHIKV rRT-PCR was clinically compared to reference molecular diagnostics for DENV - CHIKV using 182 serum samples from suspected cases in Managua, Nicaragua. The pan-DENV-CHIKV rRT-PCR had a dynamic range extending from 7.0 to 2.0 log10copies/?L for each DENV serotype - CHIKV, - the lower limits of 95% detection were 7.9-37.4copies/?L. The pan-DENV-CHIKV rRT-PCR detected DENV in 81 patients compared to 75 using a reference, hemi-nested DENV RT-PCR, - it demonstrated perfect agreement with a reference CHIKV rRT-PCR (54 positive samples). The single-reaction, multiplex format of the pan-DENV-CHIKV rRT-PCR, combined with sensitive detection of both viruses, has the potential to improve detection while decreasing testing costs - streamlining molecular workflow.
Palavras-Chave: Chikungunya virus; Dengue virus; Molecular diagnostics; Multiplex real-time RT-PCR
Imprenta: Journal of Clinical Virology, v. 78, p. 57-61, 2016
Identificador do objeto digital: 10.1016/j.jcv.2016.01.007
Descritores: Chikungunya virus - Cytopathology ; Chikungunya virus - Genome ; Chikungunya virus - Infectious diseases ; Chikungunya virus - Viral infections ; Chikungunya virus - PCR detection ; Chikungunya virus - Real Time PCR ; Chikungunya virus - RT-PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Dengue ; Chikungunya virus - Public health
Data de publicação: 2016
