Dengue Virus and Yellow Fever Virus Detection Using Reverse Transcription–Insulated Isothermal PCR and Comparison with Real-Time RT-PCR

ABSTRACT Yellow fever (YF) is a reemerging public health threat, with frequent outbreaks prompting large vaccination campaigns in regions of endemicity in Africa and South America. Specific detection of vaccine-related adverse events is resource-intensive, time-consuming, and difficult to achieve during an outbreak. To address this, we have developed a highly transferable rapid yellow fever virus (YFV) vaccine-specific real-time reverse transcription-PCR (RT-PCR) assay that distinguishes vaccine from wild-type lineages. The assay utilizes a specific hydrolysis probe that includes locked nucleic acids to enhance specific discrimination of the YFV17D vaccine strain genome. Promisingly, sensitivity and specificity analyses reveal this assay to be highly specific to vaccine strain(s) when tested on clinical samples and YFV cell culture isolates of global origin. Taken together, our data suggest the utility of this assay for use in laboratories of varied capacity for the identification and differentiation of vaccine-related adverse events from wild-type infections of both African and South American origin.

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Simultaneous detection of Dengue virus, Chikungunya virus, Zika virus, Yellow fever virus and West Nile virus

Journal of Virological Methods ◽

10.1016/j.jviromet.2019.03.014 ◽

2019 ◽

Vol 268 ◽

pp. 53-55 ◽

Cited By ~ 3

Author(s):

José A. Boga ◽

Marta E. Alvarez-Arguelles ◽

Susana Rojo-Alba ◽

Mercedes Rodríguez ◽

María de Oña ◽

...

Keyword(s):

West Nile Virus ◽

Dengue Virus ◽

Yellow Fever ◽

Zika Virus ◽

Chikungunya Virus ◽

Yellow Fever Virus ◽

Simultaneous Detection ◽

Fever Virus ◽

West Nile ◽

Virus Zika

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Yellow Fever Virus/Dengue-2 Virus and Yellow Fever Virus/Dengue-4 Virus Chimeras: Biological Characterization, Immunogenicity, and Protection against Dengue Encephalitis in the Mouse Model

Journal of Virology ◽

10.1128/jvi.77.6.3655-3668.2003 ◽

2003 ◽

Vol 77 (6) ◽

pp. 3655-3668 ◽

Cited By ~ 33

Author(s):

Thomas J. Chambers ◽

Yan Liang ◽

Deborah A. Droll ◽

Jacob J. Schlesinger ◽

Andrew D. Davidson ◽

...

Keyword(s):

Mouse Model ◽

Dengue Virus ◽

Yellow Fever ◽

Yellow Fever Virus ◽

Fever Virus ◽

E Proteins ◽

Dengue Viruses ◽

Intracerebral Inoculation ◽

Intraperitoneal Route ◽

Chimeric Viruses

ABSTRACT Two yellow fever virus (YFV)/dengue virus chimeras which encode the prM and E proteins of either dengue virus serotype 2 (dengue-2 virus) or dengue-4 virus within the genome of the YFV 17D strain (YF5.2iv infectious clone) were constructed and characterized for their properties in cell culture and as experimental vaccines in mice. The prM and E proteins appeared to be properly processed and glycosylated, and in plaque reduction neutralization tests and other assays of antigenic specificity, the E proteins exhibited profiles which resembled those of the homologous dengue virus serotypes. Both chimeric viruses replicated in cell lines of vertebrate and mosquito origin to levels comparable to those of homologous dengue viruses but less efficiently than the YF5.2iv parent. YFV/dengue-4 virus, but not YFV/dengue-2 virus, was neurovirulent for 3-week-old mice by intracerebral inoculation; however, both viruses were attenuated when administered by the intraperitoneal route in mice of that age. Single-dose inoculation of either chimeric virus at a dose of 105 PFU by the intraperitoneal route induced detectable levels of neutralizing antibodies against the homologous dengue virus strains. Mice which had been immunized in this manner were fully protected from challenge with homologous neurovirulent dengue viruses by intracerebral inoculation compared to unimmunized mice. Protection was associated with significant increases in geometric mean titers of neutralizing antibody compared to those for unimmunized mice. These data indicate that YFV/dengue virus chimeras elicit antibodies which represent protective memory responses in the mouse model of dengue encephalitis. The levels of neurovirulence and immunogenicity of the chimeric viruses in mice correlate with the degree of adaptation of the dengue virus strain to mice. This study supports ongoing investigations concerning the use of this technology for development of a live attenuated viral vaccine against dengue viruses.

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Fever versus fever: The role of host and vector susceptibility and interspecific competition in shaping the current and future distributions of the sylvatic cycles of dengue virus and yellow fever virus

Infection Genetics and Evolution ◽

10.1016/j.meegid.2013.03.008 ◽

2013 ◽

Vol 19 ◽

pp. 292-311 ◽

Cited By ~ 98

Author(s):

Kathryn A. Hanley ◽

Thomas P. Monath ◽

Scott C. Weaver ◽

Shannan L. Rossi ◽

Rebecca L. Richman ◽

...

Keyword(s):

Dengue Virus ◽

Interspecific Competition ◽

Yellow Fever ◽

Yellow Fever Virus ◽

Fever Virus

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Dengue virus detection in Aedes aegypti larvae and pupae collected in rural areas of Anapoima, Cundinamarca, Colombia

Biomédica ◽

10.7705/biomedica.v37i0.3584 ◽

2017 ◽

Vol 37 ◽

pp. 193 ◽

Cited By ~ 5

Author(s):

Myriam Lucía Velandia-Romero ◽

Víctor Alberto Olano ◽

Carolina Coronel-Ruiz ◽

Laura Cabezas ◽

María Angélica Calderón-Peláez ◽

...

Keyword(s):

Aedes Aegypti ◽

Dengue Virus ◽

Reverse Transcription ◽

Rural Areas ◽

Virus Detection ◽

Rt Pcr ◽

Transmisión Vertical

Introducción. La incidencia y la prevalencia del dengue en Cundinamarca son elevadas y, recientemente, se detectó Aedes aegypti en algunas áreas rurales del departamento.Objetivo. Evaluar la transmisión transovárica del virus del dengue en larvas y pupas recolectadas en áreas rurales del municipio de Anapoima.Materiales y métodos. Se recolectaron ejemplares vivos en 53 viviendas y se transportaron al laboratorio de Anapoima, donde se clasificaron, se agruparon y se congelaron. Llevadas a Bogotá, se las homogeneizó, se les extrajo el ARN con Trizol®, se las sometió a una reacción en cadena de la polimerasa de transcripción inversa (Reverse Transcription Polymerase Change Reaction, RT-PCR) y a PCR convencional, y los productos amplificados se analizaron en geles de agarosa al 2 %.Resultados. En 54,7 % de las viviendas evaluadas se encontraron formas inmaduras del vector y el serotipo más frecuente fue el DENV-1. Sin embargo, en algunos pools se detectó la presencia simultánea de los serotipos DENV 1 y 2, DENV 1 y 3, y DENV 1 y 4, así como los serotipos DENV 1, 2 y 3.Conclusión. Los resultados confirmaron la transmisión vertical del virus de manera natural en el área rural del municipio, lo cual reafirma la capacidad vectorial de A. aegypti y explica, en parte, la persistencia del virus en la región y la posibilidad de que en la fase adulta el vector lo transmita sin haber consumido sangre infectada. Esta situación aumenta el riesgo de infección por el virus del dengue en Colombia y, por lo tanto, la necesidad de adelantar programas de prevención y control en todas las zonas con presencia del mosquito.

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High Fidelity of Yellow Fever Virus RNA Polymerase

Journal of Virology ◽

10.1128/jvi.78.2.1032-1038.2004 ◽

2004 ◽

Vol 78 (2) ◽

pp. 1032-1038 ◽

Cited By ~ 82

Author(s):

Konstantin V. Pugachev ◽

Farshad Guirakhoo ◽

Simeon W. Ocran ◽

Fred Mitchell ◽

Megan Parsons ◽

...

Keyword(s):

Dengue Virus ◽

Rna Polymerase ◽

Yellow Fever ◽

Error Rate ◽

Yellow Fever Virus ◽

Protein A ◽

Fever Virus ◽

Genome Replication ◽

Virus Rna

ABSTRACT Three consecutive plaque purifications of four chimeric yellow fever virus-dengue virus (ChimeriVax-DEN) vaccine candidates against dengue virus types 1 to 4 were performed. The genome of each candidate was sequenced by the consensus approach after plaque purification and additional passages in cell culture. Our data suggest that the nucleotide sequence error rate for SP6 RNA polymerase used in the in vitro transcription step to initiate virus replication was as high as 1.34 × 10−4 per copied nucleotide and that the error rate of the yellow fever virus RNA polymerase employed by the chimeras for genome replication in infected cells was as low as 1.9 × 10−7 to 2.3 × 10−7. Clustering of beneficial mutations that accumulated after multiple virus passages suggests that the N-terminal part of the prM protein, a specific site in the middle of the E protein, and the NS4B protein may be essential for nucleocapsid-envelope interaction during flavivirus assembly.

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