Abstract:
:In many countries phytosanitary regulations apply to Potato spindle tuber viroid, because it can cause serious diseases in potato and tomato crops. Other pospiviroids, some of which are distributed widely in ornamental crops, can cause similar diseases. Consequently, there is a need for a reliable and cost-effective generic testing method. An assay was developed that detects all known species of the genus Pospiviroid, using real-time RT-PCR based on TaqMan technology. This GenPospi assay consists of two reactions running in parallel, the first targeting all pospiviroids, except Columnea latent viroid, the second specifically targeting the latter viroid (already published). To monitor the RNA extraction a nad5 internal control was included. Method validation on tomato leaves showed that the GenPospi assay detects all pospiviroids up to a relative infection rate of 0.13% (equals 770 times dilution). The assay was specific because no cross reactivity was observed with other viroids, viruses or nucleic acid from plant hosts. Repeatability and reproducibility were 100% and the assay appeared robust in an inter-laboratory comparison. The GenPospi assay has been shown to be a suitable tool for large-scale screening for all known pospiviroids. Although it has been validated for tomato leaves it can potentially be used for any crop.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Botermans M,van de Vossenberg BT,Verhoeven JT,Roenhorst JW,Hooftman M,Dekter R,Meekes ETdoi
10.1016/j.jviromet.2012.09.004subject
Has Abstractpub_date
2013-01-01 00:00:00pages
43-50issue
1eissn
0166-0934issn
1879-0984pii
S0166-0934(12)00307-2journal_volume
187pub_type
杂志文章abstract::A method for quantitative analysis of the growth properties of human cytomegalovirus (HCMV) in various cell culture systems was developed. Recent HCMV isolates are, in most cases cell associated, causing only limited cytopathic effect. This renders comparative analysis of the biological properties of such isolates dif...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(97)02082-x
更新日期:1997-01-01 00:00:00
abstract::In the current study, primers described previously and modified versions of these primers were evaluated for amplification of full-length gag genes from different equine infectious anemia virus (EIAV) strains from several countries, including the USA, Germany and Japan. Each strain was inoculated into a primary horse ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2012.12.010
更新日期:2013-04-01 00:00:00
abstract::Dengue surveillance relies on reverse transcription-polymerase chain reaction (RT-PCR), for confirmation of dengue virus (DENV) serotypes. We compared efficacies of published and modified primer sets targeting envelope (Env) and capsid-premembrane (C-prM) genes for detection of circulating DENV serotypes in southern I...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2017.02.014
更新日期:2017-06-01 00:00:00
abstract::Gene transfer into hepatocytes is highly desirable for the long-term goal of replacing deficient proteins and correcting metabolic disorders. Vectors based on herpes simplex virus type-1 (HSV-1) have been demonstrated to mediate efficient gene transfer into hepatocytes both in vitro and in vivo. Large transgene capaci...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2004.09.007
更新日期:2005-01-01 00:00:00
abstract::In this study, we generated recombinant virus-like particles (VLPs) against family Filoviridae, genus Ebolavirus, species Zaire ebolavirus, strain Makona (EBOV) in Drosophila melanogaster Schneider 2 (S2) cells using the EBOV Makona. S2 cells were cotransfected with four viral plasmids encoding EBOV Makona proteins an...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2018.08.016
更新日期:2018-11-01 00:00:00
abstract::A method for rapid and reliable detection of Tomato spotted wilt virus (TSWV) (Tospovirus, Bunyaviridae) in its vector Frankliniella occidentalis (Thysanoptera Thripidae) would be a useful tool for studying the epidemiology of this virus. A RT-PCR method developed for this purpose is reported. The method was tested on...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(03)00048-x
更新日期:2003-04-01 00:00:00
abstract::Beak and feather disease virus (BFDV) threatens a wide range of endangered psittacine birds worldwide. In this study, we assessed a novel PCR assay and genetic screening method using high-resolution melt (HRM) curve analysis for BFDV targeting the capsid (Cap) gene (HRM-Cap) alongside conventional PCR detection as wel...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2016.08.015
更新日期:2016-11-01 00:00:00
abstract::HSV-1-based vectors have been widely used to achieve targeted delivery of genes into the nervous system. In the current study, we aim to use shRNA-containing HSV-1-based gene delivery system for the therapy of HSV-2 infection. Guinea pigs were infected intravaginally with HSV-2 and scored daily for 100 days for the se...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.06.037
更新日期:2013-11-01 00:00:00
abstract::In situ detection of specific cells offers a unique perspective on the spatial interactions between host immune cells and specific viral pathogens or cancers. Most immunohistochemistry techniques require manual cell counting on biopsied and fixed tissue sections. The availability of sophisticated software packages for...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2015.06.004
更新日期:2015-09-15 00:00:00
abstract::Real-time PCR assays for nucleic acid testing (NAT) of hepatitis viruses A-E and for HIV-1 and HIV-2 have been developed; however, a multiplex assay that can simultaneously detect all of these agents is not yet available. Standardized TaqMan assays for detection of hepatitis viruses A-E have been described and applied...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2018.06.001
更新日期:2018-09-01 00:00:00
abstract::Hepatitis A virus (HAV) infection is the leading worldwide cause of acute viral hepatitis. An important aspect of viral control is rapid diagnosis. Epidemiological studies have linked hepatitis A outbreaks to the consumption of drinking water or soft fruits exposed to faecal contamination. Real-time reverse transcript...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2010.02.017
更新日期:2010-06-01 00:00:00
abstract::A rapid and effective ELISA for measuring Epstein-Barr virus (EBV)-neutralizing antibodies in human sera was devised to replace the existing cumbersome method involving the inhibition of fetal cord blood B-cell transformation by the virus. The new method will be invaluable for assessing antibody responses in human sub...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(98)00054-8
更新日期:1998-07-01 00:00:00
abstract::Two mosquito-borne flaviviruses, Murray Valley encephalitis (MVE) and Kunjin (KUN), are the aetiological agents of Australian encephalitis. MVE causes a severe and potentially fatal form of the disease while KUN is responsible for only a few relatively mild cases. Therefore it is important that serological tests used ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(94)00105-p
更新日期:1995-02-01 00:00:00
abstract::Human herpesviruses 6 (HHV-6), 7 (HHV-7) and 8 (HHV-8) are lymphotropic herpesviruses that may cause opportunistic diseases in immunosuppressed patients such as transplant or AIDS patients. The new commercial CMV HHV-6, 7, 8 R-gene kit (Argene, Varilhes, France) for the simultaneous quantitation of HHV-6 and qualitati...
journal_title:Journal of virological methods
pub_type: 杂志文章,多中心研究
doi:10.1016/j.jviromet.2008.01.026
更新日期:2008-05-01 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::Adequate treatment of influenza requires identification of viral type as well as detection of mutation(s) conferring drug resistance. Reverse hybridization-based line probe assays (LiPA) can be performed using several probes immobilized on nitrocellulose, strips enabling LiPA to determine simultaneously viral subtypes...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2012.06.018
更新日期:2012-11-01 00:00:00
abstract::An efficient procedure for the detection of avian reovirus (ARV)-specific RNA sequences in veterinary immunological medicinal products using reverse transcriptase polymerase chain reaction (RT-PCR) is described. Four ARV vaccine strains (1133, 1733, 2408 and Olson WVU2937), two ATCC strains (VR826 and VR856) as well a...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2004.09.019
更新日期:2005-02-01 00:00:00
abstract::Representatives from several different virus families (Baculoviridae, Herpesviridae, Myoviridae, Phycodnaviridae, Picornaviridae, Podoviridae, Retroviridae, and Siphoviridae) were stained using a variety of highly fluorescent nucleic acid specific dyes (SYBR Green I, SYBR Green II, OliGreen, PicoGreen) and examined us...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(99)00167-6
更新日期:2000-03-01 00:00:00
abstract::Two enzyme-linked immunosorbent assays for detection of adenovirus antigen in faecal extracts have been established. A conjugate of rabbit anti-(human) adenovirus immunoglobulin and horseradish peroxidase (HRP) prepared by means of a hetero-bifunctional reagent, N-succinimidyl-3-(2-pyridyldithio) propionate (SPDP), wa...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(86)90007-8
更新日期:1986-08-01 00:00:00
abstract::Disease caused by Chikungunya virus (CHIKV) is clinically characterized by sudden-onset of fever and severe arthralgia, which may persist for weeks, months, or years after acute phase of the infection. CHIKV is spreading globally; in India it first appeared in the 1960s followed by a quiescent period and then a full-b...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2016.05.003
更新日期:2016-09-01 00:00:00
abstract::Bluetongue (BT) is an arboviral disease, which can often be fatal in naïve sheep and white tailed deer, but is usually less severe, or unapparent in other ruminants. Twenty-six bluetongue virus (BTV) serotypes have been recognised so far, two of which (BTV-25 and BTV-26) were recently identified by phylogenetic compar...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.11.012
更新日期:2015-03-01 00:00:00
abstract::Specific cell-surface binding is the essential first step for cellular invasion by viruses. To understand this process, various methods to evaluate binding properties of viruses to cells have been developed. However, many rely on radioactive labeling or indirect immunofluorescence. The development of a novel fluoresce...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.08.011
更新日期:2006-06-01 00:00:00
abstract::The use of 0.1% aqueous solution of polylysine (poly-L-lysine) is proposed as a prior step to negative staining of viral or particle suspensions. Particles spread better on films precoated with polylysine than with other substances used for the same purpose. This applies particularly to samples from sucrose or CsCl gr...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(85)90121-1
更新日期:1985-05-01 00:00:00
abstract::Lymphoblastoid B.95-8 cells were cultured for four months and three weeks in the presence of increasing concentrations (50--200 microgram/ml) of phosphonoacetic acid (PAA). Several weeks after removal of the PAA, the cultures, in parallel with untreated B.95-8 cells, were tested for the presence of: 1) Epstein-Barr vi...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(80)90052-x
更新日期:1980-01-01 00:00:00
abstract::Previous evidence implicating Paramyxoviruses in the aetiopathology of Paget's disease of bone has proved controversial. Whilst several groups have demonstrated Paramyxoviruses using techniques such as in situ hybridisation (ISH), reverse transcriptase-polymerase chain reaction (RT-PCR), and in situ-RT-PCR (IS-RT-PCR)...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(03)00079-x
更新日期:2003-05-01 00:00:00
abstract::Bovine leukemia virus (BLV) is widely distributed in U.S. cattle herds. It infects B lymphocytes and causes neoplastic disease in 5-10% of infected animals. Direct economic losses are incurred as a result of death, reduced milk production and condemnation at slaughter. Thus the identification of cattle infected with B...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(91)90009-o
更新日期:1991-06-01 00:00:00
abstract::Mungbean yellow mosaic India virus (MYMIV) infecting soybean and other legumes causes yellow mosaic disease (YMD). Evaluation of soybean genotypes for YMD resistance involves field screening at disease hot spots or in a protected environment using infectious clones or viruliferous whiteflies as sources of virus inocul...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2019.03.002
更新日期:2019-06-01 00:00:00
abstract::Three anti-H5 influenza virus monoclonal antibody (mAb) clones, IFH5-26, IFH5-115 and IFH5-136, were obtained by immunising a BALB/C mouse with inactivated A/duck/Hokkaido/Vac-1/04 (H5N1). These mAbs were found to recognise specifically the haemagglutinin (HA) epitope of the influenza H5 subtypes by western blotting w...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.02.007
更新日期:2011-05-01 00:00:00
abstract::A technique for coupling foot-and-mouth disease virus (FMDV) to tanned sheep red blood cells (SRBC) is reported. Different parameters influencing the procedure were studied. Subtypes C2, C3, O1 and A24 were used as antigens, and guinea pig hyperimmune sera obtained were tested for specific antibody in passive hemagglu...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(88)90131-0
更新日期:1988-08-01 00:00:00
abstract::Nipah virus, a zoonotic paramyxovirus which emerged recently was chemically inactivated using binary ethylenimine (BEI). The inactivated virus was concentrated and purified by sucrose gradient centrifugation. The gradient fractions were examined by electron microscopy and Western immunoblot, and gradient fraction cont...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.09.005
更新日期:2006-03-01 00:00:00