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 using a standard flow cytometer equipped with a standard 15 mW argon-ion laser. The highest green fluorescence intensities were obtained using SYBR Green I. DNA viruses with genome sizes between 48.5 and 300 kb could easily be detected. The fluorescence signals of the small genome-sized RNA viruses (7.4-14.5 kb) were found at the limit of detection. No significant linear relationship could be found between genome size and the green fluorescence intensity of the SYBR Green I stained virus preparations. To our knowledge, this is the first report of detecting and discriminating between a wide range of different viruses directly using flow cytometry. This rapid and precise assay represents a new and promising tool in the field of virology.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Brussaard CP,Marie D,Bratbak Gdoi
10.1016/s0166-0934(99)00167-6subject
Has Abstractpub_date
2000-03-01 00:00:00pages
175-82issue
1-2eissn
0166-0934issn
1879-0984pii
S0166-0934(99)00167-6journal_volume
85pub_type
杂志文章abstract：:The mosaic disease caused by fig mosaic virus (FMV) is considered the plague of fig worldwide. A naïve phage display library, raised against the recombinant nucleocapsid protein of FMV (FMV-Np) was screened to obtain specific monoclonal recombinant antibodies in the form of single chain variable fragments (scFvs). Aft...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2019.113796
更新日期：2020-02-01 00:00:00
abstract：:Bubaline herpesvirus 1 (BuHV1) is a member of ruminant alphaherpesviruses antigenically related to bovine herpesvirus 1 (BoHV1). The impact of BuHV1 infection in infectious bovine rhinotracheitis control program is difficult to establish, due to the lack of specific diagnostic test. The ectodomain of glycoprotein E of...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.06.023
更新日期：2014-10-01 00:00:00
abstract：:The dot-immunobinding assay (DIBA) was adapted for detection of citrus tristeza virus (CTV) and compared with DAS-ELISA and DAS-indirect ELISA. DIBA was easy to perform and as sensitive as either ELISA procedure for CTV diagnosis. The entire test could be performed in 2-3 h using polyclonal antibodies, with minimal la...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(91)90108-c
更新日期：1991-10-01 00:00:00
abstract：:A nanoparticle-based immunoassay was developed for the rapid and sensitive detection of avian influenza virus (AIV). In this method, AIV-specific pentabody (pVHH3B) was conjugated to magnetic nanoparticles (MNPs) and used to capture AIV. Gold nanoparticles (GNPs), labelled with the anti-AIV mouse monoclonal antibody 3...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.07.024
更新日期：2010-11-01 00:00:00
abstract：:Monitoring the serum hepatitis B viral DNA levels with sensitive realtime PCR assays is strongly recommended for the management of patients with chronic HBV infection. This study compares the performance of two realtime HBV quantitative PCR assays with samples from chronic HBeAg(+) and HBeAg(-) patients. ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.06.036
更新日期：2013-11-01 00:00:00
abstract：:A novel and rapid EBNA-1 titration method has been developed which uses immunoprecipitation of specific DNA-protein complexes with EBNA-1-positive serum. The method is more sensitive than the conventional immunofluorescence method and has potential value as a diagnostic reagent for clinical laboratories. TPA induction...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(88)90062-6
更新日期：1988-09-01 00:00:00
abstract：:A polymerase chain reaction (PCR) assay was developed for detection of bovine retrospumavirus (bovine syncytial virus; BSV) provirus DNA. Two different sets of oligonucleotide primers complementary to sequences located in the gag and the pol/env gene regions were used and compared for their ability to amplify the targ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00179-7
更新日期：1999-03-01 00:00:00
abstract：:We have cloned and sequenced the glycoprotein B genes from five strains of BaCMV, isolated from three subspecies of cynocephalus baboons (olive, yellow and chacma). Primers were designed using conserved DNA regions of the gB gene to allow DNA amplification from all strains of BaCMV. These regions differ sufficiently f...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.01.010
更新日期：2005-05-01 00:00:00
abstract：:Early and accurate diagnosis of Crimean-Congo hemorrhagic fever (CCHF) is essential for the treatment and outcome of the disease and prevention of its further transmission. Molecular-based diagnostic assays now serve as the front-line tool in the diagnosis of CCHF. However, the development of real-time RT-PCR assay fo...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.11.006
更新日期：2006-05-01 00:00:00
abstract：:Quantitative PCR (qPCR) assay using TaqMan™ probe was widely used in the detection of different nucleic acids. However, this technology has several drawbacks, including false negative results caused by primer-dimer (PD) and false positive issues due to primer-probe aggregations. Here, we designed a modified TaqMan™-Mo...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.01.008
更新日期：2018-04-01 00:00:00
abstract：:Since there are indications of an increasing amount of non-B subtypes in Western Europe it was decided to assess the performance of the ViroSeq HIV-1 Genotyping System on a set of samples from the AIDS Reference Laboratory at the University Hospitals Leuven, a hospital with an increasing number of patients infected wi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2004.02.005
更新日期：2004-07-01 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：:The nucleocapsid (N) protein of a coronavirus plays a crucial role in virus assembly and in its RNA transcription. It is important to characterize a virus at the nucleotide level to discover the virus's genomic sequence variations and similarities relative to other viruses that could have an impact on the functions of...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2019.113806
更新日期：2020-03-01 00:00:00
abstract：:Loop mediated isothermal amplification (LAMP) assay is used for rapid diagnosis of Cyprinid herpesvirus-3, formerly designated koi herpesvirus (KHV), with comparable sensitivity to PCR. To reduce the time required for the LAMP assay, an immunocapture (IC) and direct binding (DB) techniques were developed to exclude th...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.07.021
更新日期：2009-12-01 00:00:00
abstract：:A solid phase reverse passive hemadsorption test (SP-RPHAd) for hepatitis B surface antigen detection is described. It was compared with a commercial reverse passive hemagglutination assay (Hepatest, Wellcome, U.K.). SP-RPHAd is four-fold less expensive than Hepatest and undiluted sera can be used instead of eight-fol...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90115-7
更新日期：1987-11-01 00:00:00
abstract：:We describe an in-situ RT-PCR method for the amplification of rhinovirus (RV) in fixed, paraffin-embedded HeLa cells employed as a model for human respiratory epithelium. HeLa cells were infected in-vitro with inocula of rhinovirus-16 ranging from 10(2) to 10(6) 50% tissue culture infective doses (TCID50), incubated f...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00095-5
更新日期：1997-09-01 00:00:00
abstract：:A real-time RT-PCR assay using SYBR Green was developed for specific and reliable quantitative detection of Citrus tristeza virus (CTV) in infected plants. A general primer set designed from conserved sequences in ORFs 1b and 2 enabled amplification of the genomic RNA (gRNA) while excluding most subgenomic and defecti...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2007.05.011
更新日期：2007-11-01 00:00:00
abstract：:A competitive reverse transcription-polymerase chain reaction (RT-PCR) was developed to detect and to quantitate the RNA of group A rotaviruses. In the assay, a 433 bp fragment is amplified by a one-tube RT-PCR protocol using primers with binding sites located in a highly conserved region of segment 6 of the rotavirus...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(02)00118-0
更新日期：2002-09-01 00:00:00
abstract：:H5N1 influenza A virus causes a rapidly fatal systemic disease in domestic poultry and spreads directly from poultry to mammalian species such as leopards, tigers and humans. The aim of this study was to develop a multiplex real-time RT-PCR for rapid detection of H5N1 influenza A virus. The selected primers and variou...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.08.004
更新日期：2006-02-01 00:00:00
abstract：:Rabies is an infectious viral disease, characterized as a neglected zoonosis, responsible for nearly 60,000 deaths annually. The virus is transmitted mainly by dogs in Africa and Asia, and wildlife in Europe and the Americas, to all mammals' species, causing severe encephalitis almost always fatal after the onset of n...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2019.04.025
更新日期：2019-08-01 00:00:00
abstract：:A transfer vector was constructed in which the neomycin resistance (neo) gene was under the control of a copy of Autographa californica nuclear polyhedrosis virus (AcMNPV) IE1 gene promoter at the p10 locus. After cotransfection of Spodoptera frugiperda (Sf9) cells with the transfer vector and infectious AcMNPV DNA, t...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(96)02112-x
更新日期：1996-10-01 00:00:00
abstract：:Adaptation through fixation of spontaneous mutations in the viral genome is considered to be one of the important factors that enable recurrent West Nile virus (WNV) outbreaks in the U.S. Genetic variations can alter viral phenotype and virulence, and degrade the performance of diagnostic and screening assays, vaccine...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.09.015
更新日期：2008-12-01 00:00:00
abstract：BACKGROUND:The HIV/HCV/HBsAg Triplex consists in manually performed, visually interpreted, lateral flow, immunochromatographic rapid diagnostic test simultaneously detecting in 15min human immunodeficiency virus (HIV)-1 and HIV-2 and hepatitis C virus (HCV)- specific antibodies (Ab) (IgG and IgM) and hepatitis B virus ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2017.12.001
更新日期：2018-03-01 00:00:00
abstract：:Commercially marketed kits are now available for rapid viral assay of clinical specimens. This study was conducted to determine the suitability of these kits for use in environmental testing. Eight rotavirus kits and one enteric adenovirus kit were screened for sensitivity using simian rotavirus SA11, human rotavirus ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90098-c
更新日期：1993-12-15 00:00:00
abstract：:Rotaviruses and non-cultivable enteric adenoviruses were purified from large volumes of faecal extracts using polyethylene glycol precipitation, centrifugation through 45% w/v sucrose and segregation by density in a cesium chloride density gradient. Yields were high and the preparations were pure enough to use in the ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(82)90059-3
更新日期：1982-08-01 00:00:00
abstract：:We have developed a murine monoclonal antibody reactive with a major immediate early 72,000 dalton protein of human cytomegalovirus and utilized this reagent in a rapid virus titration and microneutralization assay. Because of the early expression of this virus encoded protein, both assays could be accomplished within...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(89)90129-8
更新日期：1989-02-01 00:00:00
abstract：:The goal of this study was to adapt a long RT-PCR technique to amplify large PCR fragments from the genome of hepatitis C virus (HCV) isolates using clinical samples. This was done by using a reverse transcriptase devoid of RNase H activity and a mixture of two antibody-bound thermostable polymerases to combine the hi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.01.031
更新日期：2005-06-01 00:00:00
abstract：:Hepatitis B-virucidal testing of biocides in quantitative suspension tests using duck hepatitis B virus (DHBV) requires primary duck embryonic hepatocytes for viral propagation. To improve the test system and availability of these cells, commercial culture plates with different growth surfaces were tested for cell cul...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.12.004
更新日期：2009-04-01 00:00:00
abstract：:The amplification of the 468 bp fragment of the BHV-1 genome by PCR is described. The 22 bp oligomers from the BHV-1 gI gene were used as primers. For successful amplification the thermal denaturation (100 degrees C/8 min, ice) of the DNA sample was carried out prior to the cycling (95 degrees C for 1 min, 56 degrees ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90132-b
更新日期：1993-02-01 00:00:00
abstract：:We show that the immuno-gold technique adapted to electron microscopy is a sensitive method for in situ localization of viral proteins in plant cells. Using antisera raised against cauliflower mosaic virus and the viroplasm major protein (VmP) we obtained a selective labelling of the viral particles and of the viropla...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(84)90053-3
更新日期：1984-12-01 00:00:00
