Generation and characterization of Japanese encephalitis virus expressing GFP reporter gene for high throughput drug screening

Zhang, Zherui; Zhang, Hongqing; Li, Xiaodan; Deng, Cheng-Lin; Wang, Zhen; Li, Jiaqi; Li, Na; Zhang, Qiuyan; Zhang, Honglei; Zhang, Bo; Ye, Han-Qing

doi:10.1016/j.antiviral.2020.104884

Cited by 32 publications

(35 citation statements)

References 48 publications

(53 reference statements)

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“…The availability of such assays with no cross-reactivity could be of great benefit for differential serology to accurately and rapidly identify a specific flavivirus infection which is especially challenging in regions where several flaviviruses co-circulate. Finally, similar reporter assays could be optimized towards high-throughput antiviral screening campaigns, either as single (Li et al, 2020) (Zhang et al, 2020) or, by choice of compatible fluoroprotein reporters, multiplexed antiviral screens possibly allowing to identify pan-flavivirus antivirals.…”

Section: Discussionmentioning

confidence: 99%

A high-throughput neutralization assay for yellow fever serodiagnostics

Rasulova

Vercruysse

Paulissen

et al. 2021

Preprint

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Quick and accurate detection of neutralizing antibodies (nAbs) against yellow fever is essential in serodiagnosis during outbreaks, for surveillance and to evaluate vaccine efficacy in population-wide studies. All this requires serological assays that can process a large number of samples in a highly standardized format. Albeit being laborious, time-consuming and limited in throughput, classical plaque reduction neutralization test (PRNT) is still considered gold standard for the detection and quantification of nAbs due to its sensitivity and specificity. Here we report the development of an alternative fluorescence-based serological assay (SNTFLUO) with an equally high sensitivity and specificity that is fit for high-throughput testing with the potential for automation. Finally, our novel SNTFLUO was cross-validated in several reference laboratories and against international WHO standards showing its potential to be implemented in clinical use. SNTFLUO assays with similar performance are available for the Japanese encephalitis, Zika and dengue viruses amenable for differential diagnostics.IMPORTANCEFast and accurate detection of neutralizing antibodies (nAbs) against yellow fever virus (YFV) is key in yellow fever serodiagnosis, outbreak surveillance and monitoring of vaccine efficacy. Although classical PRNT still remains gold standard for measuring YFV nAbs, this methodology suffers from inherent limitations such as a low throughput and an overall high labor intensity. We present a novel fluorescence-based serum neutralization test (SNTFLUO) with equally high sensitivity and specificity that is fit for processing large number of samples in a highly standardized manner and has the potential to be implemented in clinical use. In addition, we present SNTFLUO assays with similar performance for Japanese encephalitis, Zika and dengue viruses opening new avenues for differential diagnostics.

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Section: Discussionmentioning

confidence: 99%

A high-throughput neutralization assay for yellow fever serodiagnostics

Rasulova

Vercruysse

Paulissen

et al. 2021

Preprint

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“…It is a relatively recent advancement in screening drug libraries and is highly efficient due to its accuracy in labeling the viral component. Immunofluorescent staining or construction of JEV virus expressing GFP reporter gene which generates fluorescent signal can be utilized for this assay [123,162,163]. Immunofluorescent staining involves the use of a JEV specific primary antibody (anti-JEV prM) and fluorescent-labeled secondary antibody (DyLight 488-labeled antibody).…”

Section: High Content Imagingmentioning

confidence: 99%

“…The number of cells emitting fluorescence signals are then read by high-content imaging platforms such as Cell Voyager 7000S, PerkinElmer high-content screening system, and Operetta high-content imaging system. The recorded readings are then examined using analytical software (Harmony 3.5, GraphPad Prism 5.0) to quantify each drug molecule's infectivity and effectiveness [123,162,163].…”

Section: High Content Imagingmentioning

confidence: 99%

Antiviral drug research for Japanese encephalitis: an updated review

et al. 2022

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Japanese encephalitis (JE) caused by the Japanese encephalitis virus (JEV) is one of Asia's most common viral encephalitis. JEV is a flavivirus, common in rural and sub-urban regions of Asian countries. Although only 1% of JEV-infected individuals develop JE, there is a 20–30% chance of death among these individuals and possible neurological sequelae post-infection. No licensed anti-JE drugs are currently available, despite extensive efforts to develop them. Literature search was performed using databases such as PubMed Central, Google Scholar, Wiley Online Library, etc. using keywords such as Japanese encephalitis virus, antiviral drugs, antiviral drug screening, antiviral drug targets, etc. From around 230 papers/abstracts and research reviews retrieved and reviewed for this study, approximately 180 most relevant and important ones have been cited. Different approaches in drug testing and various antiviral drug targets explored so far have been thoroughly searched from the literature and compiled, besides addressing the future perspectives of the antiviral drug development strategies. Although the development of effective anti-JE drugs is an urgent issue, only supportive care is currently available. Recent advancements in understanding the biology of infection and new drug targets have been promising improvements. Despite hindrances such as the unavailability of a proper drug delivery system or a treatment regimen irrespective of the stage of infection, several promising anti-JE candidate molecules are in different phases of clinical trials. Nonetheless, efficient therapy against JEV is expected to be achieved with drug combinations and a highly targeted drug delivery system soon. Graphical abstract

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“…In recent years, the rapid development of high-throughput screening methods has led to the discovery of a plethora of small-molecule compounds that can inhibit arboviral infection and/or replication in vertebrate cells [ 16 , 17 , 18 , 19 , 20 , 21 , 22 ], and a few also proved to be effective in mosquito cells. These anti-arboviral compounds can impede the circulation of arboviruses between arthropods and humans by either rendering arboviruses more vulnerable in humans or directly inhibiting viral infection and replication in mosquitoes.…”

Section: Introductionmentioning

confidence: 99%

Antiviral Compounds for Blocking Arboviral Transmission in Mosquitoes

Dong

Dimopoulos

2021

Viruses

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Mosquito-borne arthropod-borne viruses (arboviruses) such as the dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) are important human pathogens that are responsible for significant global morbidity and mortality. The recent emergence and re-emergence of mosquito-borne viral diseases (MBVDs) highlight the urgent need for safe and effective vaccines, therapeutics, and vector-control approaches to prevent MBVD outbreaks. In nature, arboviruses circulate between vertebrate hosts and arthropod vectors; therefore, disrupting the virus lifecycle in mosquitoes is a major approach for combating MBVDs. Several strategies were proposed to render mosquitoes that are refractory to arboviral infection, for example, those involving the generation of genetically modified mosquitoes or infection with the symbiotic bacterium Wolbachia. Due to the recent development of high-throughput screening methods, an increasing number of drugs with inhibitory effects on mosquito-borne arboviruses in mammalian cells were identified. These antivirals are useful resources that can impede the circulation of arboviruses between arthropods and humans by either rendering viruses more vulnerable in humans or suppressing viral infection by reducing the expression of host factors in mosquitoes. In this review, we summarize recent advances in small-molecule antiarboviral drugs in mammalian and mosquito cells, and discuss how to use these antivirals to block the transmission of MBVDs.

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Generation and characterization of Japanese encephalitis virus expressing GFP reporter gene for high throughput drug screening

Cited by 32 publications

References 48 publications

A high-throughput neutralization assay for yellow fever serodiagnostics

A high-throughput neutralization assay for yellow fever serodiagnostics

Antiviral drug research for Japanese encephalitis: an updated review

Antiviral Compounds for Blocking Arboviral Transmission in Mosquitoes

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