Abstract:
:A quantitative real-time PCR assay was developed for human herpesvirus-6 (HHV-6) genome based on TaqMan technology. After choosing a region of interest into the U65-U66 genes of HHV-6 genome, its nucleotide sequence was determined among four HHV-6 strains (one variant A and three variants B) to exclude a variability of sensitivity due to interstrain sequence differences. A plasmid containing HHV-6 target sequences identical to those of reference type viruses was constructed with the aim of standardisation. This HHV-6 genomic quantitation assay has a threshold sensitivity of ten copy equivalents (EqCop) per reaction. In order to test the feasibility of this assay directly on human samples, the technique was applied to the quantitation of HHV-6 genome in 30 blood samples from healthy subjects as well as 31 blood samples and three samples of cerebrospinal fluid (CSF) from 21 bone marrow transplant (BMT) recipients and four patients with a haematological disease but not treated by bone marrow transplantation. HHV-6 load ranged between 0.00015 and 0.0008 equivalent DNA copy number (EqCop) per 100 peripheral blood mononuclear cells (PBMCs) in healthy subjects whereas it ranged from <10 to 7500 EqCop per 100 PBMCs, and from <10 to 415,820 EqCop per 100 microl of whole CSF in patients. The efficacy of treatment with antiherpetic drug was associated with a decrease of the viral load in the CSF of one patient. This method leads to relevant results in term of range of quantitation, sensitivity, and safety against contamination by amplicons, and might constitute a useful tool for the follow-up of BMT recipients particularly in the presence of antiherpetic therapy.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Gautheret-Dejean A,Manichanh C,Thien-Ah-Koon F,Fillet AM,Mangeney N,Vidaud M,Dhedin N,Vernant JP,Agut Hdoi
10.1016/s0166-0934(01)00390-1subject
Has Abstractpub_date
2002-02-01 00:00:00pages
27-35issue
1-2eissn
0166-0934issn
1879-0984pii
S0166093401003901journal_volume
100pub_type
杂志文章abstract::A duplex polymerase chain reaction (PCR) was developed to measure Marek's disease virus (MDV) load in two subpopulations of Marek's disease (MD) lymphoma cells from chickens. PCR primers were designed using the sequence of the MDV-ICP4 gene and the chicken IFNgamma gene. Each set of primers was present in the same rea...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(99)00075-0
更新日期:1999-09-01 00:00:00
abstract::Recently, immunoblots (IBs) have tended to substitute Western blots (WBs) for HIV infection diagnosis. Several studies have confirmed IBs' high sensitivity to confirm HIV infection for every stage. Since the nature and pattern of the antigens of IBs are different from those of WB, the abilities of IBs and WBs to disti...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2021.114074
更新日期:2021-01-21 00:00:00
abstract::A simple and rapid solid-phase reverse transcriptase assay was developed based on the use of poly(rA):oligo(dT)12-18 as template primer immobilized on DEAE cellulose paper squares to detect human immunodeficiency virus (HIV) and/or other retroviruses in cell culture supernatants. It was found that PEG (per se) -up to ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(90)90095-w
更新日期:1990-03-01 00:00:00
abstract::The geographical distribution of human immunodeficiency virus type 1 (HIV-1) subtypes show, with the exception of some African Countries, a specific pattern. However, the significant phenomenon of migration to Western Countries, coupled to inter-ethnic blending, may result in a constant introduction and spread of nove...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2004.11.014
更新日期:2005-03-01 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 fluorogenic reverse transcriptase-polymerase chain reaction (RT-PCR) system was developed for use as a rapid diagnostic test for determining dengue viremia. The dengue virus 3'-noncoding sequence was utilized to formulate serotype-specific RT-PCR assays for quantitative identification of the four different dengue vi...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(01)00280-4
更新日期:2001-06-01 00:00:00
abstract::Herpes simplex virus (HSV) isolates were differentiated by immunoblotting analysis using a mixture of polyclonal antisera directed against HSV type 1 (HSV-1) and HSV type 2 (HSV-2) glycoprotein fractions (gB/gC of HSV-1 and gC/gE/84-kDa protein of HSV-2), since the mixed antisera recognized viral polypeptides with dif...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(87)90122-4
更新日期:1987-11-01 00:00:00
abstract::An inducible in vitro cell culture system was developed to assay HCV replication by direct biochemical means. A transcription plasmid containing a T7 promoter at the 5' end, full-length cDNA of the HCV genome, a ribozyme sequence from the antigenomic strand of hepatitis delta virus and a T7 terminator was prepared. To...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(01)00278-6
更新日期:2001-05-01 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::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::Since protection against African horsesickness (AHS) is serotype-specific, rapid serotyping of AHSV is crucial to identify the correct vaccine serotype for efficient control of the spread of AHS outbreaks, especially when they occur in non-endemic regions. This paper describes the first one-day serotyping procedure th...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2004.08.002
更新日期:2004-12-01 00:00:00
abstract::Conditions for using slot-blot nucleic acid hybridization to quantitatively detect dengue-2 virus using a radiolabelled cDNA probe, pVV17, were determined. As little as 11 plaque-forming units of virus were detected using a hybridization mixture without formamide and performing the test at 70 degrees C. While predomin...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(87)90097-8
更新日期:1987-02-01 00:00:00
abstract::Two adsorption-elution concentration methods, both involving negatively charged membranes, were evaluated in order to monitor hepatitis A virus (HAV) contamination in tap, river, mineral and coastal water samples: elution with urea-arginine phosphate buffer/reconcentration with magnesium chloride (method 1); and sodiu...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.06.008
更新日期:2006-11-01 00:00:00
abstract::Bacterial ghosts offer a new avenue for the study of inactivated vaccines. However, for many years the mechanism of genetic inactivation was controversial. To obtain mouse monoclonal antibodies (mAbs) against protein E will allow the observation of its location and dynamic expression to expand understanding of the lys...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.03.007
更新日期:2013-05-01 00:00:00
abstract::beta-Papillomaviruses (PV) seem to be involved in the pathogenesis of cutaneous squamous cell carcinoma and its early stage actinic keratosis. In this study, typing was extended of a previously described consensus primer-mediated beta- and gamma-cutaneous HPV PCR method followed by reverse-line-blotting (BGC-PCR/RLB) ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2007.05.022
更新日期:2007-12-01 00:00:00
abstract::Matching serum and oral fluid (saliva) samples were collected from 369 subjects in Tunisia in 2002, from a city in the north and a rural district in the south. Rubella-specific IgG was detected in sera by commercial ELISA (Dade Behring) and in matching oral fluids by two methods, a previously described IgG-capture ELI...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2003.09.008
更新日期:2003-12-01 00:00:00
abstract::A Western blot assay was developed and analyzed against the comparable standard, immunoprecipitation of (35)[S]-methionine/cysteine-labeled ovine progressive pneumonia virus (OPPV) proteins, for its ability to detect anti-OPPV antibodies using endpoint titers. Western blot analysis is 12-fold more sensitive in detecti...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.06.025
更新日期:2006-11-01 00:00:00
abstract::Introduction of amino acids substitutions in murine leukemia virus genome is a powerful method to determine the relative importance of various viral factors in pathogenesis. However, introduction of such amino acids substitution could result in viruses at a selective disadvantage, and eventual selection of revertants....
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.01.011
更新日期:2005-05-01 00:00:00
abstract::Loop-mediated isothermal amplification is a novel method for rapid amplification of DNA. It has been adopted widely for the detection of virus because of its simplicity, rapidity, and specificity. A loop-mediated isothermal amplification assay was developed for the detection of porcine parvovirus. Four primers specifi...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.10.004
更新日期:2009-02-01 00:00:00
abstract::Mungbean yellow mosaic virus-Vigna (MYMV) sequences cloned as partial dimers within the T-DNA of a binary vector were deleted at a high frequency upon conjugal mobilization from Escherichia coli into Agrobacterium tumefaciens. This deletion involving the genome-length viral DNA did not occur when the binary plasmid wa...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.06.002
更新日期:2006-10-01 00:00:00
abstract::Influenza vaccines must be revised constantly on almost a yearly basis because of the sequential mutations (antigenic drift) that occur as the virus responds to immunologic pressure. New, high yield (hy) reassortant viruses have proved essential to meet production needs for the supply of new vaccines. We have devised ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(01)00412-8
更新日期:2002-02-01 00:00:00
abstract::One of the most important steps when preparing a live attenuated vaccine is the assessment of the level of attenuation in target animals. It is costly and time consuming as it requires, on each occasion, a large number of susceptible animals and contained accommodation. This study assessed the consistency of the bovin...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.07.007
更新日期:2008-11-01 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::An immunocapture ELISA test for the diagnosis of human and animal influenza A and/or B is described. A monoclonal anti-nucleoprotein (NP) antibody was used to capture the NP antigen and the captured antigen was detected by an anti-NP polyclonal rabbit antiserum. Compared with the usual diagnostic method by cultivation...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(89)90102-x
更新日期:1989-07-01 00:00:00
abstract::Glycoprotein K (gK) is involved in membrane fusion phenomena during infectious virus production and egress and is an important determinant for neurovirulence. To assess better the in vitro and in vivo roles of gK in virus replication, a recombinant virus was constructed expressing an engineered enhanced green fluoresc...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(98)00107-4
更新日期:1998-11-01 00:00:00
abstract::The branched DNA (bDNA) assay for hepatitis B virus (Chiron Corporation Emerville, USA) was investigated by application to HBV-infected patients in Taiwan, where the B and C genotypes of hepatitis B virus are most prevalent. The study group included sera with hepatitis B surface antigen (HBsAg) and e antigen (HBeAg); ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(01)00335-4
更新日期:2001-08-01 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::One first generation assay (manufactured by Ortho, test I) and 3 second generation anti-HCV ELISAs (manufactured by Ortho, Abbott, and UBI, tests II-IV) were compared. Sera from 4 different sources were used: (1) intravenous drug-users (IVDUs, n = 50), (2) blood donors (n = 1055), (3) all clinical samples from one day...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(92)90087-t
更新日期:1992-12-01 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::Plaque assays under Sephadex or agarose overlays are described for rat coronaviruses (RCVs) grown in L2 mouse fibroblasts. A plaque assay using Sephadex was simple; however, viable plaques could not be collected for propagation, and fixation was necessary before evaluation. Plaque formation under agarose was optimized...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(93)90089-a
更新日期:1993-06-01 00:00:00