Improved detection of Zika virus <scp>RNA</scp> in human and animal specimens by a novel, highly sensitive and specific real‐time RT‐PCR assay targeting the 5′‐untranslated region of Zika virus

Chan, Jasper Fuk‐Woo; Yip, Cyril Chik-Yan; Tee, Kah-Meng; Zhu, Zheng; Tsang, Jessica Oi-Ling; Chik, Kenn Ka-Heng; Tsang, Terance Gi-Wai; Chan, Chris; Poon, Vincent Kwok-Man; Sridhar, Siddharth; Yin, Feifei; Hung, Ivan; Chau, Sandy; Zhang, Anna Jinxia; Chan, Kwok‐Hung; Yuen, Kwok‐Yung

doi:10.1111/tmi.12857

Cited by 36 publications

(34 citation statements)

References 51 publications

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“…Briefly, ZIKV-infected Huh-7, U251, and SF268 cells (MOI = 0.05) were treated with different concentrations of AR-12 or DMSO. The cell culture supernatants were then collected at 24 h post-inoculation (hpi), followed by total nucleic acid extraction and quantitative reverse transcription-PCR (qRT-PCR) as previously described (Chan et al, 2016c;Chan et al, 2017b). Additionally, U251 cells were inoculated with high MOI of ZIKV (MOI = 1) for 1 h. After the virus inoculum was removed, the cells were washed and cultured in fresh medium containing different concentrations of AR-12 or DMSO.…”

Section: Antiviral Evaluation Of Ar-12 In Cell Culturementioning

confidence: 99%

“…At 24 hpi, the supernatants were collected and subsequently applied for virus titration using both qRT-PCR and plaque assay. The half maximal inhibitory concentration (IC 50 ) was calculated using Sigma plot in an Excel add-in ED50V10 as we previously described (Chan et al, 2013a;Chan et al, 2017a;Chan et al, 2017b).…”

Section: Antiviral Evaluation Of Ar-12 In Cell Culturementioning

confidence: 99%

“…At 24 hpi, the supernatants were harvested in RLT buffer and RNA was extracted with the EZ1 RNA tissue mini kit (Qiagen, Hilden, Germany). The viral load was then quantified with qRT-PCR as we previously described (Chan et al, 2016c;Chan et al, 2017b). In the siRNA knockdown assay, transfection of siRNA on Huh-7 cells was performed using Lipofectamine ® RNAiMAX (Thermo Fisher Scientific) as we previously described with slight modifications (Chan et al, 2016a).…”

Section: Akt Inhibitor and Small Interfering Rna (Sirna) Knockdownmentioning

confidence: 99%

“…At 24 hpi, the supernatants were harvested in RLT buffer and RNA was extracted with the EZ1 RNA tissue mini kit (Qiagen). The viral RNA load was then quantified with qRT-PCR as we previously described (Chan et al, 2016c;Chan et al, 2017b).…”

Section: Akt Inhibitor and Small Interfering Rna (Sirna) Knockdownmentioning

confidence: 99%

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The celecoxib derivative kinase inhibitor AR-12 (OSU-03012) inhibits Zika virus via down-regulation of the PI3K/Akt pathway and protects Zika virus-infected A129 mice: A host-targeting treatment strategy

et al. 2018

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A B S T R A C TZika virus (ZIKV) is a human-pathogenic flavivirus that has recently emerged as a global public health threat. ZIKV infection may be associated with congenital malformations in infected fetuses and severe neurological and systemic complications in infected adults. There are currently limited treatment options for ZIKV infection. AR-12 (OSU-03012) is a celecoxib derivative cellular kinase inhibitor that has broad-spectrum antiviral activities. In this study, we investigated the antiviral activity and mechanism of AR-12 against ZIKV. We evaluated the in vitro anti-ZIKV activity of AR-12, using cell protection and virus yield reduction assays, in multiple clinically relevant cell lines, and the in vivo treatment effects of AR-12 in a lethal mouse model using type I interferon receptordeficient A129 mice. AR-12 inhibited ZIKV strains belonging to both the African and Asian/American lineages in Huh-7 and/or neuronal cells. AR12's IC 50 against ZIKV was consistently < 2 μM in these cells. ZIKV-infected A129 mice treated with intraperitoneally or orally administered AR-12 had significantly higher survival rate (50.0%-83.3% vs 0%, P < 0.05), less body weight loss, and lower blood and tissue ZIKV RNA loads than untreated control A129 mice. These anti-ZIKV effects were likely the results of down-regulation of the PI3K/Akt pathway by AR-12. Clinical trials using the clinically available and broad-spectrum AR-12 as an empirical treatment should be considered especially for patients residing in or returning from areas endemic of ZIKV and other arboviral infections who present with an acute undifferentiated febrile illness.

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Section: Antiviral Evaluation Of Ar-12 In Cell Culturementioning

confidence: 99%

Section: Antiviral Evaluation Of Ar-12 In Cell Culturementioning

confidence: 99%

Section: Akt Inhibitor and Small Interfering Rna (Sirna) Knockdownmentioning

confidence: 99%

Section: Akt Inhibitor and Small Interfering Rna (Sirna) Knockdownmentioning

confidence: 99%

See 2 more Smart Citations

The celecoxib derivative kinase inhibitor AR-12 (OSU-03012) inhibits Zika virus via down-regulation of the PI3K/Akt pathway and protects Zika virus-infected A129 mice: A host-targeting treatment strategy

et al. 2018

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“…Not surprisingly, when Zika virus emerged in South America and was found to be responsible for microcephaly, we received numerous requests for Zika virus, and many papers have been published with recognition of the EVAg as the source of virus. Five examples encompassing, i) simultaneous transmission with chikungunya virus, ii) diagnosis, iii) signal processing pathways, iv) pathogenesis in relation to foetal infections and v) virus structure, are cited here (Goertz et al, 2017;Chan et al, 2017;Zhang et al, 2016;Miner et al, 2016;Sirohi et al, 2016).…”

Section: The Evag Organizational Structurementioning

confidence: 99%

The European Virus Archive goes global: A growing resource for research

Romette¹,

Prat²,

Gould³

et al. 2018

Antiviral Research

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The European Virus Archive (EVA) was created in 2008 with funding from the FP7-EU Infrastructure Programme, in response to the need for a coordinated and readily accessible collection of viruses that could be made available to academia, public health organisations and industry. Within three years, it developed from a consortium of nine European laboratories to encompass associated partners in Africa, Russia, China, Turkey, Germany and Italy. In 2014, the H2020 Research and Innovation Framework Programme (INFRAS projects) provided support for the transformation of the EVA from a European to a global organization (EVAg). The EVAg now operates as a non-profit consortium, with 26 partners and 20 associated partners from 21 EU and non-EU countries. In this paper, we outline the structure, management and goals of the EVAg, to bring to the attention of researchers the wealth of products it can provide and to illustrate how end-users can gain access to these resources. Organisations or individuals who would like to be considered as contributors are invited to contact the EVAg coordinator, Jean-Louis Romette, at jean-louis.romette@univmed.fr.

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Thermo‐Responsive Polymer Capsules in Real‐Time One‐Step RT‐PCR for Highly Multiplex RNA Analysis

Kim

Jung

Kim

et al. 2020

Adv Healthcare Materials

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Rapid and simple detection of RNA targets is in high demand due to the growing threat of pandemic viruses. One‐step real‐time, reverse transcription–polymerase chain reaction (One‐step RT‐qPCR) using a controlled release system of thermo‐responsive materials is developed in this paper to enable high‐fidelity RNA analysis as suppressing by‐products. The nanocapsules, consisting of upper critical solution temperature (UCST) material and PCR primers, carry or release the primers depending upon the temperature. The UCST nanocapsules are introduced into hydrogel microparticles incorporated with RT primers and then the target RNA is selectively amplified in the microparticle through one‐step RT‐qPCR. Severe side products are sharply subdued by separating the PCR primers from the RT process by means of the microparticles with nanocapsules. Because the one‐step assay is now implemented in a single microparticle, multiple target RNAs can be analyzed in a simple RT‐qPCR of multiple particles. Reliable 18‐plex one‐step RT‐qPCR is successfully conducted within 30 min using single‐color fluorescent optics. This work also explains the facile fabrication processes used for the thermo‐responsive nanocapsules and hydrogel microparticles by the blending polymerization method. Extensible multiplex analysis of influenza virus demonstrates the versatile uses of this one‐step RT‐qPCR platform.

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Improved detection of Zika virus RNA in human and animal specimens by a novel, highly sensitive and specific real‐time RT‐PCR assay targeting the 5′‐untranslated region of Zika virus

Abstract: The highly sensitive and specific ZIKV-5'-UTR assay may help to improve the laboratory diagnosis of ZIKV infection.

Cited by 36 publications

References 51 publications

The celecoxib derivative kinase inhibitor AR-12 (OSU-03012) inhibits Zika virus via down-regulation of the PI3K/Akt pathway and protects Zika virus-infected A129 mice: A host-targeting treatment strategy

The celecoxib derivative kinase inhibitor AR-12 (OSU-03012) inhibits Zika virus via down-regulation of the PI3K/Akt pathway and protects Zika virus-infected A129 mice: A host-targeting treatment strategy

The European Virus Archive goes global: A growing resource for research

Thermo‐Responsive Polymer Capsules in Real‐Time One‐Step RT‐PCR for Highly Multiplex RNA Analysis

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