Abstract：
:A quantitative PCR method was developed in order to quantitate the number of copies of Pseudorabies virus (PRV) genome present in tissues from infected pigs. The method is based on the use of an internal standard that differs from the target DNA by a deletion of ten base pairs, and that is co-amplified with the target DNA. The resulting PCR products are labelled with a fluorescent primer and are then separated and detected by means of an automated sequencer. The assay was found to be specific and sensitive, allowing the detection of five copies of viral DNA among 10(6) host cells. The method was used successfully to quantitate the number of PRV DNA copies in trigeminal ganglia samples from infected pigs during the acute and the latent stages of the infection. Between 12 and 3.10(5) copies of viral genome per 10(6) neuronal cells were detected in these tissues which is consistent with data published previously.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Thiery R,Pannetier C,Rziha HJ,Jestin Adoi
10.1016/0166-0934(96)02072-1subject
Has Abstractpub_date
1996-09-01 00:00:00pages
79-87issue
1-2eissn
0166-0934issn
1879-0984pii
0166-0934(96)02072-1journal_volume
61pub_type
杂志文章abstract：:Hyperimmune rat antisera prepared against 5 recent antigenic variants of influenza A (H3N2) viruses were studied for haemagglutination inhibiting (HI) antibodies to the homologous and the heterologous viruses. The ratios of homologous to heterologous reactions varied from one animal to another in immunizations with ea...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(87)90147-9
更新日期：1987-03-01 00:00:00
abstract：:A rapid coliphage detection assay was developed, based on the phage-induced release of beta-galactosidase from cells of Escherichia coli. The assay could detect as few as five coliphage per sample without an overnight incubation period. The range of acceptable assay parameters was identified. ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00253-6
更新日期：2001-01-01 00:00:00
abstract：:A 4837-bp sequence of a newfound green turtle herpesvirus (GTHV), implicated in the etiology of green turtle fibropapilloma, was obtained from tumor tissues of a green turtle with fibropapilloma using a genomic walking method based on restriction enzyme digestion, self-ligation and inverse polymerase chain reaction (I...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00267-6
更新日期：2001-02-01 00:00:00
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 targete...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.09.022
更新日期：2014-12-15 00:00:00
abstract：:Procedures for cloning entire dengue serotype 2 virus genome in the multiple cloning site of a commercially available high copy number plasmid are described. The 10.7 kb viral RNA genome was reverse transcribed, amplified as three overlapping DNA fragments and successively ligated into pBluescript II KS, which contain...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00277-9
更新日期：2001-03-01 00:00:00
abstract：:A full glycoprotein E (gE) deletion was generated in genome of the Egyptian BoHV-1.1 Abu-Hammad strain. Integrity of the gE negative (gE(-)) mutant virus was proved by successful specific PCR amplifications of gB, gC, tk, gD, gI and gE genes along with definite immune reaction to polyclonal anti-BoHV-1 antibody in inf...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.07.041
更新日期：2013-12-01 00:00:00
abstract：:A quantitative real-time RT-PCR (Q-RT-PCR) was developed to detect and determine the amount of viral hemorrhagic septicemia virus (VHSV) in organs of experimentally infected rainbow trout. Primers and TaqMan probes targeting the glycoprotein (G) and the nucleoprotein (N) genes of the virus were designed. The efficienc...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.10.005
更新日期：2006-03-01 00:00:00
abstract：:Details of a procedure for detecting double-stranded RNA (dsRNA) in virus and viroid infected tissue extracts using serologically specific electron microscopy are given. A method for staining dsRNA, based on in situ formation of uranyl phosphate, that consistently permits the examination of dsRNA by electron microscop...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(84)90055-7
更新日期：1984-12-01 00:00:00
abstract：:All human herpesviruses cause chronic infections in which latent virus is periodically reactivated. Persistently active infections are uncommon, however, and occur exclusively in individuals whose immune systems fail to control virus multiplication and spread. This paper summarizes the management of these unusual infe...
journal_title：Journal of virological methods
pub_type： 杂志文章,评审
doi：10.1016/0166-0934(88)90076-6
更新日期：1988-09-01 00:00:00
abstract：:The rapid development of new molecular biology methods has improved infectious disease diagnosis, which is increasingly important to clinical management and public health. A wide variety of new methods which are more specific, sensitive and robust, such as combination of PCR and microarray technology, has gradually re...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2007.05.018
更新日期：2007-11-01 00:00:00
abstract：:Single-stranded conformation polymorphism (SSCP) is a technique used widely for the detection of differences in DNA sequence based on PCR technology. Developed by geneticists for the detection of mutations causing disease, it has been adopted more recently for the analysis of the quasi-species of viral genomes, such a...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(00)00279-2
更新日期：2001-04-01 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：:Two fragments, S66 and S55, of the S glycoprotein of the newly identified canine coronavirus type I (CCoV type I), were expressed in a procariotic system. The purified recombinant proteins of 350 and 366 amino acids in length, respectively, were employed to develop an enzyme-linked immunosorbent assay (ELISA) for the ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2003.12.003
更新日期：2004-04-01 00:00:00
abstract：:HIV-1 RNA viral load has become the major biological marker for disease prognosis and outcome of antiretroviral therapy in the treatment of HIV-infected individuals. The aim of this study was to compare the performance of the new CE marked NucliSens EasyQ HIV-1 assay with NucliSens HIV-1 QT assay (reference method). N...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.04.017
更新日期：2005-10-01 00:00:00
abstract：:Sabin type 3 polio vaccine virus is the most common cause of poliovaccine associated paralytic poliomyelitis. Vaccine associated paralytic poliomyelitis cases have been associated with Sabin type 3 revertants containing a single U to C substitution at bp 472 of Sabin type 3. A rapid method of identification of Sabin t...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00114-6
更新日期：1997-11-01 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 single amino acid substitution, from histidine to tyrosine at position 274 of the neuraminidase gene has converted Oseltamivir sensitive H5N1 influenza A virus into a resistant strain. Currently, Oseltamivir is being stockpiled in many countries potentially affected by the influenza A virus subtype H5N1 epidemic. To...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2006.09.007
更新日期：2007-01-01 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：:Noroviruses are the leading cause of nonbacterial gastroenteritis outbreaks in the United States, some of which are caused by the ingestion of contaminated water. Detection and genotypic characterization of noroviruses is commonly performed by reverse transcription-polymerase chain reaction (RT-PCR) followed by sequen...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.03.010
更新日期：2009-03-01 00:00:00
abstract：:To map the epitopes of VP2 protein of chicken anemia virus (CAV), VP2 was expressed as a fusion protein in Escherichia coli BL21 (DE3). The Western blot demonstrated that recombinant VP2 protein could be recognized by sera of chickens infected with CAV. Female BALB/c mice were immunized with purified recombinant VP2 p...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2007.03.016
更新日期：2007-08-01 00:00:00
abstract：:The polymerase chain reaction (PCR) is one of the most efficient techniques for measuring the viral load of HIV-infected samples. Determination of the specificity of PCR products is usually based on Southern blotting and hybridization of the amplified DNA to radioactive oligonucleotide probes specific for sequences co...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(94)90028-0
更新日期：1994-05-01 00:00:00
abstract：:A direct reverse transcription-polymerase chain reaction (RT-PCR) method for detecting the chrysanthemum stunt viroid (CSVd) and chrysanthemum chlorotic mottle viroid (CChMVd) to screen for a viroid-free chrysanthemum plant at a small plant size was established and named microtissue direct RT-PCR. A razor or syringe n...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2005.07.001
更新日期：2006-01-01 00:00:00
abstract：:The ferret is an excellent model for many human infectious diseases including influenza, SARS-CoV, henipavirus and pneumococcal infections. The ferret is also used to study cystic fibrosis and various cancers, as well as reproductive biology and physiology. However, the range of reagents available to measure the ferre...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.04.014
更新日期：2014-09-01 00:00:00
abstract：:Large-scale serosurveillance of severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) will only be possible if serological tests are sufficiently reliable, rapid and affordable. Many assays are either labour-intensive and require specialised facilities (e.g. virus neutralization assays), or are expensive w...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2020.114025
更新日期：2021-02-01 00:00:00
abstract：:A recombinant Newcastle disease virus (NDV) expressing the green fluorescent protein (GFP) was generated by applying reverse genetics techniques. The GFP open reading frame flanked by NDV transcription start and stop sequences was inserted between the fusion (F)- and hemagglutinin-neuraminidase genes in a full-length ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(02)00247-1
更新日期：2003-03-01 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：:A sensitive enzyme-linked immunosorbent assay (ELISA) has been developed which can detect serotype and group-specific antibodies (IgG) to fowl adenovirus serotypes 2, 3 and 4. The chickens produced principally type-specific antibodies after a single oral inoculation of virus which enabled that strain to be identified ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(83)90086-1
更新日期：1983-12-01 00:00:00
abstract：:An efficient method for isolation of virus mutants with antigenically altered proteins is described. The method is based on the separation of viruses with wild-type and antigenically altered proteins by affinity chromatography using monoclonal antibodies (MAbs). A nonessential glycoprotein E (gE) of Aujeszky's disease...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(98)00137-2
更新日期：1999-01-01 00:00:00
abstract：:OLIGSCAN (oligonucleotide scanner) is a computer program for IBM-PC-compatible computers that allows the user to scan up to 200 DNA sequences for homology to oligonucleotide sequences of interest. Once a core sequence of longer than the user-defined minimum length is found, the remainder of the oligonucleotide is comp...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(92)90143-2
更新日期：1992-02-01 00:00:00
abstract：:A method for assessing the number of infectious particles in preparations of HIV has been developed. Virus was mixed with cells to allow binding of virus. The cells were then cast in an agar gel to block any further transfer of virus between the cells. After 4 days of incubation the cells initially infected with HIV e...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(90)90133-z
更新日期：1990-02-01 00:00:00
