Abstract：
:We applied real-time RT-PCR to the analysis of Tobacco rattle virus (TRV)-mediated virus-induced gene silencing (VIGS) of the phytoene desaturase (PDS) gene in Nicotiana benthamiana and tomato. Using a combination of direct measurement and mathematical assessment, we evaluated three plant genes, ubiquitin (ubi3), elongation factor-1 alpha (EF-1), and actin, for use as internal reference transcripts and found that EF-1 and ubi3 were least variable under our experimental conditions. Primer sets designed to amplify the 5' or 3' regions of endogenous PDS transcripts in tomato yielded similar reductions in transcript levels indicating a uniform VIGS-mediated degradation of target RNA. By measuring the ratio of the abundance of the PDS insert transcript to the TRV coat protein RNA, we established that the PDS insert within TRV was stable in both hosts. VIGS in N. benthamiana resulted in complete photo-bleaching of all foliar tissue compared to chimeric bleaching in tomato. PDS transcript levels were decreased eleven- and seven-fold in photobleached leaves of N. benthamiana and tomato, respectively, while sampling tomato leaflets on the basis of age rather than visible bleaching resulted in only a 17% reduction in PDS coupled with a large leaf-to-leaf variation. There was a significant inverse relationship (r2=76%, P=0.01) between the relative abundance of CP RNA and the amount of PDS transcript in rTRV::tPDS-infected tomato suggesting that virus spread and accumulation are required precursors for successful VIGS in this host.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Rotenberg D,Thompson TS,German TL,Willis DKdoi
10.1016/j.jviromet.2006.07.017subject
Has Abstractpub_date
2006-12-01 00:00:00pages
49-59issue
1-2eissn
0166-0934issn
1879-0984pii
S0166-0934(06)00272-2journal_volume
138pub_type
杂志文章abstract：:The in vitro syncytium induction capacity of human immunodeficiency virus type 1 (HIV1) isolates is an important marker in the progression of the disease. Two methods have been widely used to determine the biological phenotype of HIV1. These two methods, the direct MT-2 assay and the supernatant assay, were compared f...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(96)02152-0
更新日期：1997-03-01 00:00:00
abstract：:Rice tungro disease (RTD) is a recurring disease affecting rice farming especially in the South and Southeast Asia. The disease is commonly diagnosed by visual observation of the symptoms on diseased plants in paddy fields and by polymerase chain reaction (PCR). However, visual observation is unreliable and PCR can be...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2016.12.001
更新日期：2017-02-01 00:00:00
abstract：:A simple, sensitive and specific colourimetric hybridisation method for the detection of HCMV DNA in clinical specimens is described. This method combines a PCR assay with a sensitive sandwich hybridisation assay. It relies on the use of a specific capture probe linked covalently to polystyrene microplates and a speci...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(99)00023-3
更新日期：1999-07-01 00:00:00
abstract：:In recent years, there has been an increase in reported cases of fowl adenovirus serotype 4 (FAdV-4) in chickens in China. The use of live attenuated vaccines contaminated with FAdV-4 has been proved to be one of the important causes of massive outbreaks of hydropericardium syndrome. To detect the contamination with F...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.09.005
更新日期：2019-03-01 00:00:00
abstract：:Canine distemper, caused by Canine distemper virus (CDV), is a highly contagious and fatal systemic disease in free-living and captive carnivores worldwide. Accurate, rapid and simple detection of CDV is critical to improve disease management and prevent outbreaks. In this study, a visible and incubation instrument-fr...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.07.007
更新日期：2018-10-01 00:00:00
abstract：:Real-time polymerase chain reaction (real-time PCR) is an accurate, rapid and reliable method that can be used for the detection and also for the quantitation of specific DNA molecules. It can be non-specific, with intercalating dyes (SYBR Green I dye) able to bind to any dsDNA, or specific with a probe (TaqMan), wher...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.05.010
更新日期：2012-09-01 00:00:00
abstract：:CD4 T-cell count is a priority for staging HIV disease and guiding clinical management as part of HIV care. Conventional CD4 T-cell enumeration methods based on flow cytometry are expensive, require well-trained personnel, and are challenging to use in rural, resource-scarce areas. A simple CD4 T-cell count test that ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2019.05.012
更新日期：2019-09-01 00:00:00
abstract：:Healthcare workers (HCWs) are at significantly higher risk of exposure to influenza virus during seasonal epidemics and global pandemics. During the 2009 influenza pandemic, some healthcare organizations recommended that HCWs wear respiratory protection such as filtering facepiece respirators, while others indicated t...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.05.009
更新日期：2018-10-01 00:00:00
abstract：:A mRNA in situ hybridization method was developed to detect mRNA of Epstein--Barr virus-determined nuclear antigen-2 (EBNA2). Strong in situ hybridization signals were detected in EBNA2-transfected cells with the antisense probe but not with the sense probe of this mRNA. Hybridization signals were also found in formal...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00271-8
更新日期：2001-04-01 00:00:00
abstract：:Lumpy skin disease virus (LSDV) is a poxvirus that causes systemic disease in cattle, resulting in substantial economic loss to affected communities. LSDV is a rapidly emerging pathogen of growing global concern that recently spread from Africa and the Middle East into Europe and Asia, impacting the cattle population ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2020.113943
更新日期：2020-11-01 00:00:00
abstract：:This study describes the development and evaluation of a cost effective test rationale for the detection of anti-HCV in dried blood spots. Samples were screened using an 'in house' IgG ELISA that incorporated the recombinant proteins c22-3, c200 and NS5. Confirmation of specific antibody to HCV was by a modification o...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00127-4
更新日期：1997-11-01 00:00:00
abstract：:Dengue virus (DENV) nonstructural-1 (NS1) glycoprotein is useful for diagnosis of DENV infections in the first 8 days of illness with any of the four serotypes (DENV-1, DENV-2, DENV-3 and DENV-4). However, NS1 diagnostics are less sensitive for secondary DENV infections so the utility of NS1 diagnostics in dengue ende...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.06.003
更新日期：2015-09-15 00:00:00
abstract：:The development of a rapid and highly sensitive PCR assay for the detection of human rhinoviruses (HRVs) in nasopharyngeal aspirates is described. Two simple and fast commercial RNA extraction methods and four primer pairs were compared. The most sensitive RNA extraction method (Ultraspec) and primer pair (A) were app...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00049-9
更新日期：1997-06-01 00:00:00
abstract：:Classical swine fever (CSF) is a highly contagious disease, causing severe economic losses in the pig industry worldwide. Vaccination of pigs with lapinized Chinese vaccines is still practised in some regions of the world, where the virus is enzootic, in order to prevent and control the disease. However, a single real...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.05.003
更新日期：2011-08-01 00:00:00
abstract：:Laser microdissection combined with real-time RT-PCR presents an advanced tool to quantify particular RNA species in defined tissue areas. Dealing with infectious tissue samples increases the need to overcome the risk of infectivity and contamination during laser microdissection. Here, an useful method to control infe...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2006.03.024
更新日期：2006-08-01 00:00:00
abstract：:The genes encoding the Erns and E2 antigen epitopes of classical swine fever virus (CSFV) were expressed as a chimeric protein in Escherichia coli BL21 by pET expression system. The antigenicity of the expressed protein CnC2 was identified by indirect enzyme-linked immunoabsorbant assay (ELISA) and immunoblot with ant...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.12.006
更新日期：2012-03-01 00:00:00
abstract：:Bovine viral diarrhea virus (BVDV) should be a ubiquitous viral pathogen to the cattle and sheep industry. This pathogen is responsible for severe economic losses. We previously showed that plasmid-mediated dual short hairpin RNA (shRNA) efficiently inhibit BVDV replication in bovine kidney epithelial (MDBK) cells. In...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.03.014
更新日期：2015-06-15 00:00:00
abstract：:A bovine in vitro organ culture (BIVOC) system was evaluated as a model to study host and pathogen events during the course of bovine herpesvirus-1 infection. Upper respiratory tract epithelium, from slaughtered animals, was cultured in an air-liquid interface system and integrity, viability, and TNF-alpha gene expres...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.02.001
更新日期：2009-06-01 00:00:00
abstract：:The morphogenesis and the ultrastructure of a marine fish iridovirus isolated from diseased grouper, Epinephelus tauvina were studied by electron microscopy. The virus was grown on a marine fish cell line (GP) at 25 degrees C. After appearance of advanced cytopathic effect (CPE), various morphogenetic stages of virus ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00350-0
更新日期：2001-10-01 00:00:00
abstract：:Simultaneous detection of two antigens by immunostaining usually requires primary antibodies from two different species or a hapten modification of one of the antibodies if they are from the same species. A novel double staining method is described for immunodetection of two independent antigens using two mouse monocl...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00048-2
更新日期：1998-07-01 00:00:00
abstract：:Live and inactivated preparations of foot-and-mouth disease virus strains 01 BFS 1860 and A22 IRQ 24/64 were freeze-dried in the presence or absence of additive solutions and assessed for their reactivity by ELISA at intervals over a six month storage period at various temperatures and also after reconstitution and su...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(90)90019-c
更新日期：1990-11-01 00:00:00
abstract：:Immune electron microscopy (IEM) cannot be used successfully for structures that do not have recognisable morphology. However, at least some of these structures or components are related antigenically to recognisable antigens or viruses. We have therefore mixed unknown antigens with known markers and looked for the pr...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(81)90035-5
更新日期：1981-02-01 00:00:00
abstract：:An immunocapture ELISA test for the diagnosis of human and animal influenza A and/or B is described. A monoclonal anti-nucleoprotein (NP) antibody was used to capture the NP antigen and the captured antigen was detected by an anti-NP polyclonal rabbit antiserum. Compared with the usual diagnostic method by cultivation...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(89)90102-x
更新日期：1989-07-01 00:00:00
abstract：:Certification represents the first line of defense against fruit tree viruses. For certification or surveys dealing with large number of samples, ELISA is still considered the technique of choice and requires a continuous supply of good quality antibodies. Prune dwarf virus (PDV) is among the major viruses affecting s...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2004.05.013
更新日期：2004-10-01 00:00:00
abstract：:An in situ hybridization technique has been optimised for use on paraffin-embedded sections of tissues collected from cattle infected experimentally with foot-and-mouth disease virus type O1BFS. Tissue was collected 5 days after infection by direct contact. In situ hybridization was carried out using an RNA probe corr...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(94)00153-8
更新日期：1995-01-01 00:00:00
abstract：:Dengue virus infection is a major public health problem throughout tropical countries. In endemic areas, dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS) are common complications resulting in death. However, serological confirmation of dengue-related illness is often complicated and time-consuming. Detect...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(95)00026-q
更新日期：1995-08-01 00:00:00
abstract：:Codon-optimization refers to the alteration of gene sequences, to make codon usage match the available tRNA pool within the cell/species of interest. Codon-optimization has emerged as a powerful tool to increase protein expression by genes from small RNA and DNA viruses, which commonly contain overlapping reading fram...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(03)00173-3
更新日期：2003-08-01 00:00:00
abstract：:Canine serum preserved at room temperature (25°C) for longer than 24h is known to exhibit significant cytotoxicity. This phenomenon is one of the major reasons for the failure of virus neutralization tests. In this study, a method for reducing this cytotoxicity was investigated by applying several treatments to dog, c...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.08.018
更新日期：2015-12-01 00:00:00
abstract：:The Pr55gag human immunodeficiency virus type 1 (HIV-1) precursor protein that is capable of auto-assembling was used as a carrier for a consensus sequence of the principal neutralization domain (PND) of the HIV-1 envelope. For this purpose, a modified HIV-1 gag gene with deletion of the sequence encoding a previously...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(94)00100-u
更新日期：1995-02-01 00:00:00
abstract：:Hepatitis C virus (HCV) infection is a major worldwide health problem, causing chronic hepatitis, liver cirrhosis and primary liver cancer (Hepatocellular carcinoma). HCV encodes a precursor polyprotein that is enzymatically cleaved to release the individual viral proteins. The viral non-structural proteins are cleave...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(02)00255-0
更新日期：2003-02-01 00:00:00
