Real-Time RT-PCR Assays for Detection and Genotyping of West Nile Virus Lineages Circulating in Africa

Fall, Gamou; Faye, Martin; Weidmann, Manfred; Kaiser, Marco; Dupressoír, Anne; Ndiaye, El Hadj Mamadou; Bâ, Yamar; Diallo, Mawlouth; Faye, Ousmane; Sall, Amadou Alpha

doi:10.1089/vbz.2016.1967

Cited by 18 publications

(21 citation statements)

References 34 publications

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“…RNA was extracted from cell suspensions and supernatants and copy numbers of genome were quantified by real time RT-PCR as previously described [ 98 ]. Infectious viral particles were measured in supernatants by plaque assay also as previously described [ 99 ].…”

Section: Methodsmentioning

confidence: 99%

“…For each lineage, 2.4x10 3 PFU were used for kinetics in mosquito and mammal cells. The ratio of particles per infectious unit in the initial viral stocks ranged from 8 to 600 [ 98 ]. Our viral stocks had a similar ratio of particles per infectious unit as that seen produced by fully infectious extracellular WN virus particles [ 100 ] and mosquito-derived replicon WN virus particles [ 101 ].…”

Section: Methodsmentioning

confidence: 99%

“…For cell fractions, prior to RNA extraction, cells were lysed by serial cycles of freeze/thaw. For the detection and quantification of viral RNA, a consensus WNV real-time RT-PCR assay and corresponding RNA standard were used as previously described [ 98 ]. The real-time PCR assays were performed using the Quantitect Probe RT-PCR Kit (Qiagen, Heiden, Germany) in a 96-well plate under the following conditions: 50°C for 10 min, 95°C for 15 min followed by 40 cycles of 95°C for 15 s and 60°C for 1 min.…”

Section: Methodsmentioning

confidence: 99%

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Biological and phylogenetic characteristics of West African lineages of West Nile virus

et al. 2017

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The West Nile virus (WNV), isolated in 1937, is an arbovirus (arthropod-borne virus) that infects thousands of people each year. Despite its burden on global health, little is known about the virus’ biological and evolutionary dynamics. As several lineages are endemic in West Africa, we obtained the complete polyprotein sequence from three isolates from the early 1990s, each representing a different lineage. We then investigated differences in growth behavior and pathogenicity for four distinct West African lineages in arthropod (Ap61) and primate (Vero) cell lines, and in mice. We found that genetic differences, as well as viral-host interactions, could play a role in the biological properties in different WNV isolates in vitro, such as: (i) genome replication, (ii) protein translation, (iii) particle release, and (iv) virulence. Our findings demonstrate the endemic diversity of West African WNV strains and support future investigations into (i) the nature of WNV emergence, (ii) neurological tropism, and (iii) host adaptation.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Biological and phylogenetic characteristics of West African lineages of West Nile virus

et al. 2017

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“…For other arboviruses, RT-PCR for Chikungunya [20], dengue [21], West Nile [22], yellow fever [23], Zika [24], and CCHF [25] viruses was performed as previously described. The RNA was amplified using ABI Prism 7000 SDS Real-Time apparatus (Applied Biosystems) with the QuantiTect kit (Qiagen).…”

Section: Methodsmentioning

confidence: 99%

Detection of the Northeastern African Rift Valley Fever Virus Lineage During the 2015 Outbreak in Mauritania

Bob

Fall

et al. 2017

Open Forum Infectious Diseases

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Background.Rift Valley fever (RVF) is an acute viral anthropozoonosis that causes epizootics and epidemics among livestock population and humans. Multiple emergences and reemergences of the virus have occurred in Mauritania over the last decade. This article describes the outbreak that occurred in 2015 in Mauritania and reports the results of serological and molecular investigations of blood samples collected from suspected RVF patients.Methods.An RVF outbreak was reported from 14 September to 26 November 2015 in Mauritania. Overall, 184 suspected cases from different localities were identified by 26 health facilities. Blood samples were collected and tested by enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) at the Institut Pasteur de Dakar (IPD). Sequencing of partial genomes and phylogenetic analyses were performed on RT-PCR–positive samples. As part of routine surveillance at IPD, samples were also screened for dengue, yellow fever, West Nile, Crimean Congo hemorrhagic fever, Zika, and Chikungunya viruses by ELISA and RT-PCR.Results.Of the 184 suspected cases, there were 57 confirmed cases and 12 deaths. Phylogenetic analysis of the sequences indicated an emergence of a virus that originated from Northeastern Africa. Our results show co-circulation of other arboviruses in Mauritania—dengue, Crimean Congo hemorrhagic fever, and West Nile viruses.Conclusion.The Northeastern Africa lineage of RVF was responsible for the outbreak in Mauritania in 2015. Co-circulation of multiples arboviruses was detected. This calls for systematic differential diagnosis and highlights the need to strengthen arbovirus surveillance in Africa.

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“…West Nile virus (WNV) was first isolated in Africa, West Nile district of Uganda, in 1937 [1]. WNV is a member of the Japanese encephalitis complex which also includes Japanese encephalitis virus, St. Louis encephalitis virus, Rocio virus, and Murray Valley encephalitis virus [2].…”

Section: Introductionmentioning

confidence: 99%

Seroprevalence of West Nile Virus among Arabian Horses in Central, Eastern Anatolia, and Marmara, Turkey

Akkutay-Yoldar¹,

Tezcan-Ülger²

2018

azb

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The West Nile Virus (WNV), a mosquito-borne Flavivirus is a common infection identified in the Middle East, Europe, West Asia, Africa, Australia and the United States. The virus can cause severe illnesses and death in horses. The current study aimed to screen West Nile virus neutralizing antibodies in Arabian horse populations from breeding farms located in Central, Eastern Anatolia and Marmara by plaque reduction neutralization test (PRNT). We sampled 165 Arabian racehorses, during March-December, 2010, in three different provinces (Eskişehir, Malatya, and Bursa) in Turkey. Screening of serum samples showed that 6.6% were positive for West Nile (WN) virus-neutralizing antibodies. The obtained data demonstrate WNV is circulating in mentioned regions among racehorses and the need to carry out a surveillance program includes WNV-focused screening tests in Turkey.

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Real-Time RT-PCR Assays for Detection and Genotyping of West Nile Virus Lineages Circulating in Africa

Cited by 18 publications

References 34 publications

Biological and phylogenetic characteristics of West African lineages of West Nile virus

Biological and phylogenetic characteristics of West African lineages of West Nile virus

Detection of the Northeastern African Rift Valley Fever Virus Lineage During the 2015 Outbreak in Mauritania

Seroprevalence of West Nile Virus among Arabian Horses in Central, Eastern Anatolia, and Marmara, Turkey

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