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https://www.readbyqxmd.com/read/28214373/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
#1
Jasper Fuk-Woo Chan, Cyril Chik-Yan Yip, Kah-Meng Tee, Zheng Zhu, Jessica Oi-Ling Tsang, Kenn Ka-Heng Chik, Terance Gi-Wai Tsang, Chris Chung-Sing Chan, Vincent Kwok-Man Poon, Siddharth Sridhar, Feifei Yin, Ivan Fan-Ngai Hung, Sandy Ka-Yee Chau, Anna Jinxia Zhang, Kwok-Hung Chan, Kwok-Yung Yuen
OBJECTIVE AND METHOD: We developed and evaluated five novel real-time RT-PCR assays targeting conserved regions in the 5'-untranslated region (5'-UTR), envelope (E'), non-structural protein 2A (NS2A), NS5, and 3'-UTR of the ZIKV genome. RESULTS: The ZIKV-5'-UTR assay exhibited the lowest in-vitro limit of detection (5-10 RNA copies/reaction and 3.0×10(-1) plaque-forming units/ml). Compared to the modified version of a widely adopted RT-PCR assay targeting the ZIKV-E gene, the ZIKV-5'-UTR assay showed better sensitivity in human clinical specimens, and representative mouse specimens, including many organs which are known to be involved in human ZIKV infection but difficult to obtain in clinical settings...
February 18, 2017: Tropical Medicine & International Health: TM & IH
https://www.readbyqxmd.com/read/28196656/a-cdna-clone-launched-platform-for-high-yield-production-of-inactivated-zika-vaccine
#2
Yujiao Yang, Chao Shan, Jing Zou, Antonio E Muruato, Diniz Nunes Bruno, Barbosa de Almeida Medeiros Daniele, Pedro F C Vasconcelos, Shannan L Rossi, Scott C Weaver, Xuping Xie, Pei-Yong Shi
A purified inactivated vaccine (PIV) using the Zika virus (ZIKV) Puerto Rico strain PRVABC59 showed efficacy in monkeys, and is currently in a phase I clinical trial. High-yield manufacture of this PIV is essential for its development and vaccine access. Here we report an infectious cDNA clone-launched platform to maximize its yield. A single NS1 protein substitution (K265E) was identified to increase ZIKV replication on Vero cells (a cell line approved for vaccine production) for both Cambodian FSS13025 and Puerto Rico PRVABC59 strains...
February 6, 2017: EBioMedicine
https://www.readbyqxmd.com/read/27940224/the-calmodulin-antagonist-w-7-n-6-aminohexyl-5-chloro-1-naphthalenesulfonamide-hydrochloride-inhibits-denv-infection-in-huh-7-cells
#3
Patricia Bautista-Carbajal, Ruben Soto-Acosta, Antonio H Angel-Ambrocio, Margot Cervantes-Salazar, Circe I Loranca-Vega, Mayra Herrera-Martínez, Rosa M Del Angel
Dengue virus (DENV) replicative cycle occurs in the endoplasmic reticulum where calcium ions play an important role in cell signaling. Calmodulin (CaM) is the primary sensor of intracellular Ca(2+) levels in eukaryotic cells. In this paper, the effect of the calmodulin antagonist W-7 in DENV infection in Huh-7 cells was evaluated. W7 inhibited viral yield, NS1 secretion and viral RNA and protein synthesis. Moreover, luciferase activity, encoded by a DENV replicon, was also reduced. A decrease in the replicative complexes formation was clearly observed in W7 treated cells...
January 15, 2017: Virology
https://www.readbyqxmd.com/read/27746901/illustrating-and-homology-modeling-the-proteins-of-the-zika-virus
#4
Sean Ekins, John Liebler, Bruno J Neves, Warren G Lewis, Megan Coffee, Rachelle Bienstock, Christopher Southan, Carolina H Andrade
The Zika virus (ZIKV) is a flavivirus of the family Flaviviridae, which is similar to dengue virus, yellow fever and West Nile virus. Recent outbreaks in South America, Latin America, the Caribbean and in particular Brazil have led to concern for the spread of the disease and potential to cause Guillain-Barré syndrome and microcephaly. Although ZIKV has been known of for over 60 years there is very little in the way of knowledge of the virus with few publications and no crystal structures. No antivirals have been tested against it either in vitro or in vivo...
2016: F1000Research
https://www.readbyqxmd.com/read/27667714/trim52-inhibits-japanese-encephalitis-virus-replication-by-degrading-the-viral-ns2a
#5
Wenchun Fan, Mengge Wu, Suhong Qian, Yun Zhou, Huanchun Chen, Xiangmin Li, Ping Qian
The members of tripartite-motif containing (TRIM) protein participate in various cellular processes and play an important role in host antiviral function. TRIM proteins exert their antiviral activity either directly by degrading viral proteins through their E3 ligase activity, or indirectly by promoting host innate immunity. This study demonstrated for the first time that TRIM52 is a novel antiviral TRIM protein against Japanese encephalitis virus (JEV) infection. Overexpression of TRIM52 restricted JEV replication in BHK-21 and 293T cells...
September 26, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27665292/a-simple-method-for-developing-an-infectious-cdna-clone-of-japanese-encephalitis-virus
#6
Hao Zheng, Xuchen Zheng, Wu Tong, Fei Liu, Chao Liang, Tao Wang, Fei Gao, Liwei Li, Tongling Shan, Guoxin Li, Guangzhi Tong
Flavivirus cDNA clones frequently demonstrate genetic instability in transformed bacteria, which hampers the construction and manipulation of cDNAs for infectious flaviviruses. In this study, we developed a stable, full-length cDNA clone, pJEHEN, of a GI JEV strain HEN0701 using a medium-copy-number pBR322 vector and propagating cDNA clones at room temperature. The virus vJEHEN recovered from the infectious clone was indistinguishable from the parent virus HEN0701 with respect to plaque morphology, growth kinetics, and virulence characteristics...
February 2017: Virus Genes
https://www.readbyqxmd.com/read/27590716/computational-prediction-and-analysis-of-potential-antigenic-ctl-epitopes-in-zika-virus-a-first-step-towards-vaccine-development
#7
Manas R Dikhit, Md Yousuf Ansari, Vijaymahantesh, Kalyani, Rani Mansuri, Bikash R Sahoo, Budheswar Dehury, Ajay Amit, Roshan K Topno, Ganesh C Sahoo, Vahab Ali, Sanjiva Bimal, Pradeep Das
The Zika virus disease is an Aedes mosquito-borne disease caused by the ZIKA virus. The unavailability of vaccines or proper chemotherapeutic treatment emphasizes the need for the development of preventive and therapeutic vaccines. T cell specific epitopes have been used as vaccine candidates to generate desired immune responses against a variety of viral pathogens. Herein, the immune-informatics approach was used for the screening of potential major histocompatibility complex class I restricted epitopes, which may be competent to generate a cell-mediated immune response in humans...
August 31, 2016: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/27443615/hla-drb1-alleles-are-associated-with-different-magnitudes-of-dengue-virus-specific-cd4-t-cell-responses
#8
Daniela Weiskopf, Michael A Angelo, Alba Grifoni, Patrick H O'Rourke, John Sidney, Sinu Paul, Aruna D De Silva, Elizabeth Phillips, Simon Mallal, Sunil Premawansa, Gayani Premawansa, Ananda Wijewickrama, Bjoern Peters, Alessandro Sette
BACKGROUND: Each year dengue virus (DENV) infects 400 million human but causes symptomatic disease in only a subset of patients, suggesting that host genetic factors may play a role. HLA molecules that restrict T-cell responses are one of the most polymorphic host factors in humans. METHODS: Here we map HLA DRB1-restricted DENV-specific epitopes in individuals previously exposed to DENV, to identify the breadth and specificity of CD4(+) T-cell responses. To investigate whether HLA-specific variations in the magnitude of response might predict associations between dengue outcomes and HLA-DRB1 alleles, we assembled samples from hospitalized patients with known severity of disease...
October 1, 2016: Journal of Infectious Diseases
https://www.readbyqxmd.com/read/27296393/predicting-zika-virus-structural-biology-challenges-and-opportunities-for-intervention
#9
Bryan D Cox, Richard A Stanton, Raymond F Schinazi
BACKGROUND: Zika virus is an emerging crisis as infection is implicated in severe neurological disorders-Guillain-Barré syndrome and fetal microcephaly. There are currently no treatment options available for Zika virus infection. This virus is part of the flavivirus genus and closely related to Dengue Fever Virus, West Nile Virus, and Japanese Encephalitis Virus. Like other flaviviruses, the Zika virus genome encodes three structural proteins (capsid, precursor membrane, and envelope) and seven nonstructural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5)...
August 2015: Antiviral Chemistry & Chemotherapy
https://www.readbyqxmd.com/read/27273223/first-isolation-of-west-nile-virus-from-a-dromedary-camel
#10
Sunitha Joseph, Ulrich Wernery, Jade Ll Teng, Renate Wernery, Yi Huang, Nissy Ag Patteril, Kwok-Hung Chan, Shyna K Elizabeth, Rachel Yy Fan, Susanna Kp Lau, Jörg Kinne, Patrick Cy Woo
Although antibodies against West Nile virus (WNV) have been detected in the sera of dromedaries in the Middle East, North Africa and Spain, no WNV has been isolated or amplified from dromedary or Bactrian camels. In this study, WNV was isolated from Vero cells inoculated with both nasal swab and pooled trachea/lung samples from a dromedary calf in Dubai. Complete-genome sequencing and phylogenetic analysis using the near-whole-genome polyprotein revealed that the virus belonged to lineage 1a. There was no clustering of the present WNV with other WNVs isolated in other parts of the Middle East...
June 8, 2016: Emerging Microbes & Infections
https://www.readbyqxmd.com/read/27169727/population-genomics-of-dengue-virus-serotype-4-insights-into-genetic-structure-and-evolution
#11
Vaishali P Waman, Sunitha Manjari Kasibhatla, Mohan M Kale, Urmila Kulkarni-Kale
The spread of dengue disease has become a global public health concern. Dengue is caused by dengue virus, which is a mosquito-borne arbovirus of the genus Flavivirus, family Flaviviridae. There are four dengue virus serotypes (1-4), each of which is known to trigger mild to severe disease. Dengue virus serotype 4 (DENV-4) has four genotypes and is increasingly being reported to be re-emerging in various parts of the world. Therefore, the population structure and factors shaping the evolution of DENV-4 strains across the world were studied using genome-based population genetic, phylogenetic and selection pressure analysis methods...
August 2016: Archives of Virology
https://www.readbyqxmd.com/read/27053551/transmembrane-domains-of-ns2b-contribute-to-both-viral-rna-replication-and-particle-formation-in-japanese-encephalitis-virus
#12
Xiao-Dan Li, Cheng-Lin Deng, Han-Qing Ye, Hong-Lei Zhang, Qiu-Yan Zhang, Dong-Dong Chen, Pan-Tao Zhang, Pei-Yong Shi, Zhi-Ming Yuan, Bo Zhang
UNLABELLED: Flavivirus nonstructural protein 2B (NS2B) is a transmembrane protein that functions as a cofactor for viral NS3 protease. The cytoplasmic region (amino acids 51 to 95) alone of NS2B is sufficient for NS3 protease activity, whereas the role of transmembrane domains (TMDs) remains obscure. Here, we demonstrate for the first time that flavivirus NS2B plays a critical role in virion assembly. Using Japanese encephalitis virus (JEV) as a model, we performed a systematic mutagenesis at the flavivirus conserved residues within the TMDs of NS2B...
June 15, 2016: Journal of Virology
https://www.readbyqxmd.com/read/26907325/in-vitro-and-in-vivo-evaluation-of-mutations-in-the-ns-region-of-lineage-2-west-nile-virus-associated-with-neuroinvasiveness-in-a-mammalian-model
#13
Katalin Szentpáli-Gavallér, Stephanie M Lim, László Dencső, Krisztián Bányai, Penelope Koraka, Albert D M E Osterhaus, Byron E E Martina, Tamás Bakonyi, Ádám Bálint
West Nile virus (WNV) strains may differ significantly in neuroinvasiveness in vertebrate hosts. In contrast to genetic lineage 1 WNVs, molecular determinants of pathogenic lineage 2 strains have not been experimentally confirmed so far. A full-length infectious clone of a neurovirulent WNV lineage 2 strain (578/10; Central Europe) was generated and amino acid substitutions that have been shown to attenuate lineage 1 WNVs were introduced into the nonstructural proteins (NS1 (P250L), NS2A (A30P), NS3 (P249H) NS4B (P38G, C102S, E249G))...
February 19, 2016: Viruses
https://www.readbyqxmd.com/read/26384477/the-a66g-back-mutation-in-ns2a-of-jev-sa14-14-2-strain-contributes-to-production-of-ns1-protein-and-the-secreted-ns1-can-be-used-for-diagnostic-biomarker-for-virulent-virus-infection
#14
Jingman Wang, Xinfeng Li, Jinyan Gu, Yu Fan, Peng Zhao, Ruibing Cao, Puyan Chen
Japanese encephalitis virus (JEV) is the most common cause of the prevalent encephalitis in Asia-Pacific region and poses a serious risk to public health. Here, we developed a reliable reverse genetics system based on the JEV SA14-14-2 strain to further explore the mechanism for the synthesis of NS1' protein and to investigate the function of NS1' protein during virus infection. NS1' is an additional form of NS1 protein with 52 amino acid carboxy-terminal extension and is expressed by the members of the Japanese encephalitis (JE) serogroup due to the translation frameshift...
December 2015: Infection, Genetics and Evolution
https://www.readbyqxmd.com/read/26377679/proteolytic-cleavage-analysis-at-the-murray-valley-encephalitis-virus-ns1-2a-junction
#15
COMPARATIVE STUDY
Siti Nor Khadijah Addis, Eva Lee, Jayaram Bettadapura, Mario Lobigs
BACKGROUND: Our understanding of the proteolytic processing events at the NS1-2A junction in the flavivirus polyprotein has not markedly progressed since the early work conducted on dengue virus (DENV). This work identified an octapeptide sequence located immediately upstream of the cleavage site thought to be important in substrate recognition by an as yet unknown, endoplasmic reticulum-resident host protease. Of the eight amino acid recognition sequence, the highly conserved residues at positions P1, P3, P5, P7 and P8 (with respect to N-terminus of NS2A) are particularly sensitive to amino acid substitutions in terms of DENV NS1-NS2A cleavage efficiency; however, the role of the octapeptide in efficient NS1 and NS2A production of other flaviviruses has not been experimentally addressed...
2015: Virology Journal
https://www.readbyqxmd.com/read/25968648/dengue-virus-ns-proteins-inhibit-rig-i-mavs-signaling-by-blocking-tbk1-irf3-phosphorylation-dengue-virus-serotype-1-ns4a-is-a-unique-interferon-regulating-virulence-determinant
#16
Nadine A Dalrymple, Velasco Cimica, Erich R Mackow
UNLABELLED: Dengue virus (DENV) replication is inhibited by the prior addition of type I interferon or by RIG-I agonists that elicit RIG-I/MAVS/TBK1/IRF3-dependent protective responses. DENV infection of primary human endothelial cells (ECs) results in a rapid increase in viral titer, which suggests that DENV inhibits replication-restrictive RIG-I/interferon beta (IFN-β) induction pathways within ECs. Our findings demonstrate that DENV serotype 4 (DENV4) nonstructural (NS) proteins NS2A and NS4B inhibited RIG-I-, MDA5-, MAVS-, and TBK1/IKKε-directed IFN-β transcription (>80%) but failed to inhibit IFN-β induction directed by STING or constitutively active IRF3-5D...
May 12, 2015: MBio
https://www.readbyqxmd.com/read/25915765/profiling-of-viral-proteins-expressed-from-the-genomic-rna-of-japanese-encephalitis-virus-using-a-panel-of-15-region-specific-polyclonal-rabbit-antisera-implications-for-viral-gene-expression
#17
Jin-Kyoung Kim, Jeong-Min Kim, Byung-Hak Song, Sang-Im Yun, Gil-Nam Yun, Sung-June Byun, Young-Min Lee
Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, is closely related to West Nile (WN), yellow fever (YF), and dengue (DEN) viruses. Its plus-strand genomic RNA carries a single open reading frame encoding a polyprotein that is cleaved into three structural (C, prM/M, and E) and at least seven nonstructural (NS1/NS1', NS2A, NS2B, NS3, NS4A, NS4B, and NS5) proteins, based on previous work with WNV, YFV, and DENV. Here, we aimed to profile experimentally all the viral proteins found in JEV-infected cells...
2015: PloS One
https://www.readbyqxmd.com/read/25811657/clinical-outcome-and-genetic-differences-within-a-monophyletic-dengue-virus-type-2-population
#18
Hapuarachchige Chanditha Hapuarachchi, Rachel Choon Rong Chua, Yuan Shi, Tun Lin Thein, Linda Kay Lee, Kim Sung Lee, David Chien Lye, Lee Ching Ng, Yee Sin Leo
The exact mechanisms of interplay between host and viral factors leading to severe dengue are yet to be fully understood. Even though previous studies have implicated specific genetic differences of Dengue virus (DENV) in clinical severity and virus attenuation, similar studies with large-scale, whole genome screening of monophyletic virus populations are limited. Therefore, in the present study, we compared 89 whole genomes of DENV-2 cosmopolitan clade III isolates obtained from patients diagnosed with dengue fever (DF, n = 58), dengue hemorrhagic fever (DHF, n = 30) and dengue shock syndrome (DSS, n = 1) in Singapore between July 2010 and January 2013, in order to determine the correlation of observed viral genetic differences with clinical outcomes...
2015: PloS One
https://www.readbyqxmd.com/read/25787282/a-single-amino-acid-substitution-in-the-ns2a-protein-of-japanese-encephalitis-virus-affects-virus-propagation-in-vitro-but-not-in-vivo
#19
Yuki Takamatsu, Kouichi Morita, Daisuke Hayasaka
We identified a unique amino acid of NS2A113, phenylalanine, that affects the efficient propagation of two Japanese encephalitis virus strains, JaTH160 and JaOArS982, in neuroblastoma Neuro-2a cells but not in cell lines of extraneural origin. This amino acid did not affect viral loads in the brain or survival curves in mice. These findings suggest that virus propagation in vitro may not reflect the level of virus neuroinvasiveness in vivo.
June 2015: Journal of Virology
https://www.readbyqxmd.com/read/25694595/a-basic-cluster-in-the-n-terminus-of-yellow-fever-virus-ns2a-contributes-to-infectious-particle-production
#20
Stephanie Voßmann, Janett Wieseler, Romy Kerber, Beate Mareike Kümmerer
UNLABELLED: The flavivirus NS2A protein is involved in the assembly of infectious particles. To further understand its role in this process, a charged-to-alanine scanning analysis was performed on NS2A encoded by an infectious cDNA clone of yellow fever virus (YFV). Fifteen mutants containing single, double, or triple charged-to-alanine changes were tested. Five of them did not produce infectious particles, whereas efficient RNA replication was detectable for two of the five NS2A mutants (R22A-K23A-R24A and R99A-E100A-R101A mutants)...
May 2015: Journal of Virology
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