Abstract：
:A recombinant murine cytomegalovirus (mCMV) that expresses enhanced green fluorescent protein (EGFP) under control of the native immediate-early 1/3 promoter was constructed to detect directly sites of viral activity in latent and reactivated infections. The recombinant virus had acute and latent infection characteristics similar to those of wild-type mCMV. Rare green-fluorescing foci were observed in paraffin sections from lungs and spleens infected latently. Positive immunoperoxidase staining for EGFP in sections of the same lung tissues suggests that these cells may be sites of restricted viral gene expression. EGFP was detected easily in tissue explants reactivating from latent infection in vitro. Morphology and adhesion characteristics of fluorescing cells suggest that viral reactivation occurs in tissue macrophages in explant cultures. The observations presented in this study demonstrate the usefulness of EGFP-expressing recombinants as tools for direct tracking of mCMV activity in vivo and in vitro.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Henry SC,Schmader K,Brown TT,Miller SE,Howell DN,Daley GG,Hamilton JDdoi
10.1016/s0166-0934(00)00202-0subject
Has Abstractpub_date
2000-09-01 00:00:00pages
61-73issue
1-2eissn
0166-0934issn
1879-0984pii
S0166093400002020journal_volume
89pub_type
杂志文章abstract：:In the context of early infant diagnosis (EID) decentralization in sub-Saharan Africa, dried blood spot (DBS) is now widely used for HIV proviral DNA detection in resource-limited settings. A new version of CAP/CTM (version 2) has been introduced, recently by Roche Diagnosis as a new real-time PCR assay to replace pre...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.12.006
更新日期：2016-03-01 00:00:00
abstract：:Tissue culture ID50 and plaque assays for the Autographa californica nuclear polyhedrosis virus, using the Spodoptera littoralis cell line HPB-SL, were developed. Direct comparison using these assay methods showed that these cells were as susceptible to infection as the more commonly used Spodoptera frugiperda cell li...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(85)90082-5
更新日期：1985-01-01 00:00:00
abstract：:A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of abalone herpesvirus DNA. Two pairs of primers were designed, based on the sequence of the DNA polymerase gene of abalone herpesvirus. The reaction temperature and time were optimized to 63°C and 60min, respectively. LAMP amplicons...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.11.011
更新日期：2014-02-01 00:00:00
abstract：:The genus Parapoxvirus includes four members, bovine papular stomatitis virus (BPSV), pseudocowpox virus (PCPV), orf virus (ORFV) and parapoxvirus of red deer in New Zealand (PVNZ). A set of primers for polymerase chain reaction (PCR) was designed to detect viral DNA from cells infected with each of the four parapoxvi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(99)00144-5
更新日期：2000-02-01 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：:Recently, sequences from a putative member of the Flaviviridae, GB virus C (GBV-C), were isolated from the serum of patients with cryptogenic hepatitis. These sequences were 83-99% identical at the nucleotide level. Because of the divergence between these GBV-C isolates, it is likely that the PCR-based detection assay...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(95)01956-1
更新日期：1996-01-01 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：:Peach rosette mosaic disease was first described in the 1940s affecting peach and plum. It was later determined that peach rosette mosaic virus (PRMV) is the causal agent of the disease. PRMV, a member of the genus Nepovirus, infects several perennial crops including stone fruit, grape and blueberry as well as several...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.01.004
更新日期：2018-04-01 00:00:00
abstract：:The aim of this study was to evaluate viral DNA load in paediatric patients with herpes simplex infections of the central nervous system. A real-time PCR assay for herpes simplex types 1 and 2 was developed for this 8-year retrospective study that included children with herpes simplex infection of the central nervous ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2007.09.009
更新日期：2008-02-01 00:00:00
abstract：:The species cowpox virus (CPXV), genus Orthopoxvirus (OPV), consists of isolates highly variable in their biological properties and their genotypes. A TaqMan PCR assay for the specific detection of CPXV DNA based on sequences of the ORF D11L has been developed recently. (Gavrilova et al., 2010; Shchelkunov et al., 201...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.10.004
更新日期：2015-01-01 00:00:00
abstract：:A multiplex polymerase chain reaction (multiplex-PCR) to detect and differentiate bovine herpesvirus 1 (BoHV-1) and 5 (BoHV-5) was developed using primers for the gene sequence that encodes the glycoprotein C. The technique was assessed against the BoHV-1 and BoHV-5 cell culture adapted strains, and clinical samples c...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.05.001
更新日期：2005-09-01 00:00:00
abstract：:Foot-and-mouth disease is one of the most economically important virus diseases of livestock. Two important requirements for the control of this disease are rapid laboratory diagnosis and epidemiological investigation. The use of the polymerase chain reaction method (PCR) to amplify specific nucleic acid regions offer...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90176-r
更新日期：1993-04-01 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：:A microtiter cell-culture assay is described for measuring neutralizing antibody activity directed against the human immunodeficiency virus (HIV). The assay relies upon inhibition of HIV-mediated cell killing of infected MT-4 lymphoid cells. The assay exhibits comparable sensitivity to two other methods used for such ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(88)90123-1
更新日期：1988-07-01 00:00:00
abstract：:5-min boiling of 3-fold diluted serum leads to the quantitative release of immune-complexed HIV antigen (Ag) that can be readily measured. The method is capable of detecting as little as 4 to 5 pg/ml Ag p24 depending on the Ag test used. Of a panel of 50 anti-HIV-positive sera, 30 (60%) were Ag-positive after heat den...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90080-b
更新日期：1993-07-01 00:00:00
abstract：:Infection with high-risk types of human papillomavirus (HPV) is necessary for the development of cervical cancer. However, the majority of the HPV infections are efficiently cleared by the immune system and only a minority persist and induce the development of malignant lesions. Several studies provided evidence that ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2004.02.018
更新日期：2004-08-01 00:00:00
abstract：:A dot-immunobinding assay (DIBA) was optimized and used successfully for the rapid detection of 15 known viruses [Alfalfa mosaic virus (AMV), Bean pod mottle virus (BPMV), Bean yellow mosaic virus (BYMV), Cowpea mild mottle virus (CPMMV), Cowpea severe mosaic virus (CPSMV), Cucumber mosaic virus (CMV), Peanut mottle v...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2017.09.003
更新日期：2017-11-01 00:00:00
abstract：:Porcine reproductive and respiratory syndrome virus (PRRSV) became a significant pathogen of swine upon its emergence in the late 1980s and since then has exemplified a rapidly evolving, constantly re-emerging pathogen. In addition to the challenges faced in development of vaccines and diagnostics, research on the bas...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.02.007
更新日期：2015-06-15 00:00:00
abstract：:Congenital human cytomegalovirus infections are the major infectious cause of birth defects in the United States. How this virus crosses the placenta and causes fetal disease is poorly understood. Guinea pig cytomegalovirus (GPCMV) is a related virus that provides an important model for studying cytomegaloviral congen...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.01.031
更新日期：2008-05-01 00:00:00
abstract：:Infection with human papillomavirus (HPV) is an important etiological factor in the development of cervical cancer, and detection of the viral genome is of prognostic importance, particularly for preneoplastic lesions. We developed a simple, easy and efficient non-organic method of DNA extraction from cervical scrapes...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(92)90157-9
更新日期：1992-01-01 00:00:00
abstract：:The recent emergence of novel viruses requires reliable methodology for their identification and confirmation both on a cellular and molecular level. Mass spectrometry offers a suitable approach for the identification and characterisation of viral proteins and its application is demonstrated in this study. Menangle vi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00441-4
更新日期：2002-04-01 00:00:00
abstract：:The results of polymerase chain reaction (PCR) and nested polymerase chain reaction (n-PCR) assays for the diagnosis of canine coronavirus (CCV) infection, and the comparison with other diagnostic techniques, such as electron microscopy (EM) and virus isolation using A-72 cell line are reported. The study was carried ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(99)00134-2
更新日期：2000-01-01 00:00:00
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), elon...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2006.07.017
更新日期：2006-12-01 00:00:00
abstract：:Rapid alternative methods are required to evaluate easily acyclovir (ACV) sensitivity of clinical herpes simplex virus (HSV) isolates. The objective of this study was to screen 54 ACV-sensitive and 41 ACV-resistant clinical HSV-1 isolates, well characterized by phenotypic and genotypic methods, for the phosphorylation...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.12.001
更新日期：2013-03-01 00:00:00
abstract：:Recently, polyomaviruses KI and WU were identified in the airways of patients with acute respiratory symptoms. The epidemiology and pathogenesis of these two viruses are not fully understood, and the development of molecular assays, such as Real Time PCR, was useful for examining their biology and role in different cl...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.07.016
更新日期：2009-12-01 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：:RT-qPCR, an established method for the detection of RNA viruses, requires internal RNA controls for the correct interpretation of PCR results. Robust and versatile RT-PCR controls can be achieved for example by packaging RNA into a virus-derived protein shell. In this study a MS2-based internal control system was deve...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.07.028
更新日期：2014-11-01 00:00:00
abstract：:Photobiotin was used to prepare biotinylated, nonradioactive nucleic acid probes for the detection of the RNA of barley yellow dwarf virus (BYDV) in plant extracts. A 1.7-kb cDNA of the PAV isolate of BYDV in the plasmid pUC8 vector was biotinylated and then used intact or as sonicated double-stranded DNA fragments. S...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90007-3
更新日期：1987-06-01 00:00:00
abstract：:The recently developed semi-automatic Hepatube system was evaluated in comparison to another radioimmunoassay for the detection of hepatitis B surface antigen (HBsAg), the manual Ausria II-125 test. After incubation of serum in anti-HBs coated tubes, the Hepatube system uses a machine to wash the tubes and to add trac...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(83)90074-5
更新日期：1983-02-01 00:00:00
abstract：:Ultra-deep pyrosequencing (UDPS) of targeted amplicons allows to determine a large number of individual sequence reads from a single PCR product, and this approach is thus extremely valuable for analysis of mixed viral populations. A mixture of genetically distinct DNA templates, however, may lead to the generation of...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.07.040
更新日期：2010-10-01 00:00:00
