Abstract:
:Accurate detection and quantitation of viruses can be beneficial to plant-virus interaction studies. In this study, three SYBR green real-time RT-PCR assays were developed to quantitate grapevine leafroll-associated virus 3 (GLRaV-3) in infected vines. Three genomic regions (ORF1a, coat protein and 3'UTR) were targeted to quantitate GLRaV-3 relative to three stably expressed reference genes (actin, GAPDH and α-tubulin). These assays were able to detect all known variant groups of GLRaV-3, including the divergent group VI, with equal efficiency. No link could be established between the concentration ratios of the different genomic regions and subgenomic RNA (sgRNA) expression. However, a significant lower virus concentration ratio for plants infected with variant group VI compared to variant group II was observed for the ORF1a, coat protein and the 3'UTR. Significant higher accumulation of the virus in the growth tip was also detected for both variant groups. The quantitation of viral genomic regions under different conditions can contribute to elucidating disease aetiology and enhance knowledge about virus ecology.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Bester R,Pepler PT,Burger JT,Maree HJdoi
10.1016/j.jviromet.2014.09.022subject
Has Abstractpub_date
2014-12-15 00:00:00pages
67-75eissn
0166-0934issn
1879-0984pii
S0166-0934(14)00377-2journal_volume
210pub_type
杂志文章abstract::Avian influenza virus (AIV) is an enveloped virus with segmented RNA that belongs to the Orthomyxoviridae. Recently, avian influenza virus isolates have not only posed a significant threat to the poultry industry but also serious public health concerns. The full-length viral hemagglutinin (HA), neuraminidase (NA) or b...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.05.006
更新日期:2011-08-01 00:00:00
abstract::Genetic reassortment of influenza viruses is widely used for creating viruses with specific phenotypes. Reassortment of two influenza viruses, each with eight RNA segments potentially yields as many as 256 gene segment combinations. Therefore, confirmation that progeny viruses possess genomes corresponding to the spec...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2004.11.024
更新日期:2005-03-01 00:00:00
abstract::Arenaviruses merit significant attention both as causative agents of endemic hemorrhagic fevers and as model systems to study the immune response to acute and persistent viral infections. Development of highly sensitive quantitative screening methods to detect arenavirus is critical for early diagnosis of patients, to...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2009.04.001
更新日期:2009-08-01 00:00:00
abstract::E(rns) is an envelope glycoprotein of classical swine fever virus (CSFV) with RNase activity. The purpose of this study was to produce an active E(rns) for further applications using the yeast secreted expression system. The E(rns) gene was cloned into the expression vector pGAPZalphaC which was introduced into Pichia...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.08.020
更新日期:2006-03-01 00:00:00
abstract::The aim of this study was to develop a more specific and sensitive competitive inhibition ELISA (CI-ELISA) than the currently used indirect ELISA for detection of antibodies to bovine immunodeficiency virus (BIV) in cattle and buffaloes. Murine monoclonal antibodies (MAbs) were generated against a recombinant capsid (...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.11.008
更新日期:2008-03-01 00:00:00
abstract::Arboviruses in the genus Flavivirus are major causes of human disease worldwide, and yet they are widely under reported. This is partially due to the classical methods of detection, using virologic or immunologic techniques, which typically lack sensitivity and specificity or require biological containment facilities ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.05.001
更新日期:2007-10-01 00:00:00
abstract::A RT-nested PCR that amplifies part of the conserved nucleoprotein gene of avian Paramyxovirus type 1 is described. The technique allowed the detection of pigeon Paramyxovirus type 1 (pPMV-1) virus directly from a wide range of infected chicken and pigeon organs, and should be able to detect typical Newcastle disease ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(02)00148-9
更新日期:2002-12-01 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::A specific padlock probe (PLP) and detection probe were designed to target the capsid protein gene of Taura syndrome virus (TSV), and a hyper-branched rolling circle amplification (HRCA) assay and a corresponding strip-based test were produced. The reaction time and temperature were optimized for both. The PLPs were l...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.08.019
更新日期:2014-12-01 00:00:00
abstract::TaqMan Mismatch Amplification Mutation Assay (TaqMAMA) is a highly sensitive allelic discrimination method. The mismatch amplification mutation assay (MAMA) is based on preferential amplification of mutant allele by the 'MAMA' primer, which is designed to have two mismatches with the wild-type allele and only one mism...
journal_title:Journal of virological methods
pub_type: 杂志文章,随机对照试验
doi:10.1016/j.jviromet.2008.07.020
更新日期:2008-11-01 00:00:00
abstract::Two genotype-specific fluorogenic RT-PCR assays were developed for the detection and quantitation of canine coronavirus (CCoV) type I and type II RNA in the faeces of dogs with diarrhoea. Both the fluorogenic assays showed high specificity, sensitivity and reproducibility, allowing a precise quantitation of CCoV type ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.06.005
更新日期:2005-12-01 00:00:00
abstract::Measles virus (MV) enters cells by binding to the signaling lymphocyte activation molecule (also called CD150) on the cell surface, and thus shows the lymphotropism and immunosuppressive effects. The head domain (residues Asp149 to Arg617) of the MV hemagglutinin (MV-H), the attachment protein, was produced using a tr...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.01.008
更新日期:2008-04-01 00:00:00
abstract::Human cosaviruses (HCoSVs) are newly discovered viruses in Picornaviridae family. Until now, most published studies reported HCoSV detection using molecular techniques and genetic characterization of the virus. Nevertheless, no laboratory has yet reported the replication of these viruses in cultured cell lines. In the...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.06.011
更新日期:2014-10-01 00:00:00
abstract::Multiplication of encephalomyocarditis virus (EMCV) in human HEp-2 cells, and its suppression by interferon (IFN), was demonstrated by direct enzyme immunoassay (EIA) in cell culture. EMCV was detected in glutaraldehyde fixed HEp-2 cell monolayers, in wells of 96-well plates, with a horse radish peroxidase (HRPO) labe...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(89)90017-7
更新日期:1989-04-01 00:00:00
abstract::Understanding the ability to survive in an aerosol leads to better understanding of the hazard posed by pathogenic organisms and can inform decisions related to the control and management of disease outbreaks. This basic survival information is sometimes lacking for high priority select agents such as the filoviruses ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.06.021
更新日期:2011-10-01 00:00:00
abstract::To formulate inactivated influenza vaccines, the concentration of hemagglutinin (HA) must be accurately determined. The standard test currently used to measure HA in influenza vaccines is the Single Radial Immunodiffusion (SRID) assay. We developed a very rapid, simple and sensitive alternative quantitative HA assay, ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2017.10.011
更新日期:2018-01-01 00:00:00
abstract::A new approach to simultaneous detection and typing of related agents by the multiplex polymerase chain reaction (PCR) is described. The reaction was been applied to human herpesviruses by nested amplification of fragments of the DNA polymerase genes. During the first amplification, primers were used as two equimolar ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(93)90061-u
更新日期:1993-10-01 00:00:00
abstract::A new, versatile baculovirus vector was developed for the generation of recombinants in the yeast Saccharomyces cerevisiae and for the expression of foreign proteins in both insect larvae and in insect cells. This vector is based on Autographa californica multiple nucleocapsid nucleopolyhedrovirus (AcMNPV) and exploit...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(97)00109-2
更新日期:1997-10-01 00:00:00
abstract::Hepatitis B micelles containing the p25 component of hepatitis B surface antigen (HBsAg) have been produced by Triton X-100 solubilization followed by ultracentrifugation in linear sucrose gradients. The product was found to resemble micelle forms prepared from plasma-derived HBsAg with the surface being composed of d...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(86)90004-2
更新日期:1986-08-01 00:00:00
abstract::Japanese encephalitis (JE) is caused by the Japanese encephalitis virus (JEV). It is a major public health problem in Asia. JEV infects swine which results in fatal encephalitis, abortion and stillbirth in pregnant sow, and hypospermia in boars. Swine is a viral amplifier, and thus plays a critical role in JEV transmi...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2010.04.015
更新日期:2010-09-01 00:00:00
abstract::Determining whether animals have been infected with foot-and-mouth disease virus or vaccinated is important because infected animals frequently become carriers of the virus, shed it intermittently and thus may be the source of new outbreaks of the disease. We had shown previously that the sera of convalescent animals ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(96)02165-9
更新日期:1997-04-01 00:00:00
abstract::A method for detecting the emergence of potential pandemic-causing influenza strains has been developed. The system first uses real-time RT-PCR to detect H5, the highly pathogenic avian influenza subtype most likely to cause a pandemic. Pyrosequencing is then employed to scan for codon changes encoding amino acids kno...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.09.013
更新日期:2008-12-01 00:00:00
abstract::Prunus necrotic ringspot ilarvirus (PNRSV) exists as a number of biologically distinct variants which differ in host specificity, serology, and pathology. Previous nucleotide sequence alignment and phylogenetic analysis of cloned reverse transcription-polymerase chain reaction (RT-PCR) products of several biologically...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(99)00051-8
更新日期:1999-07-01 00:00:00
abstract::Recombinant plasmids carrying 199 base pairs (bp) inserts from the non coding region (nucleotides (nt) 6-204) of the TT virus (TTV) genome were used to standardize an heteroduplex mobility assay able to detect mixed infections of a single individual with several TTV isolates. In this simplified heteroduplex mobility a...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(01)00425-6
更新日期:2002-03-01 00:00:00
abstract::The diagnostic system based on reverse transcription (RT)-PCR has been used widely for the detection of viral genomes of faecal-borne RNA viruses. However, faecal specimens often produce both false positive and false negative results. Therefore, there is a need for a diagnosis procedure that can control for 'false-res...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(02)00016-2
更新日期:2002-07-01 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::A one-tube real-time TaqMan reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the detection of equine arteritis virus (EAV). The test was validated using the seminal plasma and nasal secretions of infected horses that were proven to contain EAV by traditional virus isolation in rabbit ki...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(01)00416-5
更新日期:2002-03-01 00:00:00
abstract::Xenotropic murine leukemia virus (X-MuLV) is often used in retrovirus elimination studies involving rodent cells. Currently, X-MuLV is measured using a focus-forming assay on mink (MiCl1 S+L-) or cat (PG-4 S+L-) cell lines. An easier and quicker PG-4 cell plaque assay, which retains the statistical reproducibility of ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(99)00064-6
更新日期:1999-08-01 00:00:00
abstract::Cell culture based assays used to detect waterborne viruses typically call for incubating the sample for at least two weeks in order to ensure that all the culturable virus present is detected. Historically, this estimate was based, at least in part, on the length of time used for detecting poliovirus. In this study, ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2016.09.002
更新日期:2016-11-01 00:00:00
abstract::The serological diagnosis of primary postnatal rubella infection is based on detection of rubella-virus-specific IgM antibody or a four-fold rise in rubella-specific IgG antibody. Although there are several different methods of enzyme immunoassays that are commercially available, the cost benefit evaluation makes them...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(91)90119-k
更新日期:1991-09-01 00:00:00