Abstract:
:A novel multiplex nested PCR (nPCR) method was developed for detecting and differentiating simultaneously the DNA of polyomaviruses JC, BK and SV40 in a single tube. In the first amplification step the same set of primers were used to amplify a conserved DNA region of the large T antigen gene of JCV, BKV and SV40. The second round of multiplex nPCR was carried out using a set of primers designed to render products of different size for each related virus. The thermocycling parameters and concentration of each reaction component were optimised systematically to achieve optimal specificity and sensitivity for the nPCR assay. The sensitivity of the method ranged between one and 10 copies of polyomavirus genome. Cerebrospinal fluid (CSF) was examined from AIDS patients with clinical and neuroradiological evidence of progressive multifocal leukoencephalopathy (PML) and CSF from AIDS patients with other neurological alterations. Urine specimens from bone marrow transplant recipients affected by haemorrhagic cystitis were also tested. The results obtained suggest that the assay is a good tool for supporting the diagnosis of polyomavirus infection and could be used for epidemiological purposes and in other studies in order to define better the role of polyomaviruses in human disease.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Fedele CG,Ciardi M,Delia S,Echevarria JM,Tenorio Adoi
10.1016/s0166-0934(99)00095-6subject
Has Abstractpub_date
1999-10-01 00:00:00pages
137-44issue
2eissn
0166-0934issn
1879-0984pii
S0166093499000956journal_volume
82pub_type
杂志文章abstract::In 2013, a unique seventh yellow head virus genotype (YHV7) was detected in Black Tiger shrimp (Penaeus monodon) broodstock that suffered high mortality following their capture from Joseph Bonaparte Gulf (JBG) in northern Australia. To assist with its diagnosis and assessment of its distribution, prevalence and pathog...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2019.113689
更新日期:2019-11-01 00:00:00
abstract::A technique based on the reverse transcriptase-polymerase chain reaction (RT-PCR) has been developed to detect the presence of Prunus necrotic ringspot virus (PNRSV) and prune dwarf virus (PDV) simultaneously in almond. This paper presents the results of a 3-year study comparing both enzyme-linked immunosorbent assay ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2003.08.014
更新日期:2003-12-01 00:00:00
abstract::Coxsackievirus B4 (CBV-4) capsid protein VP0 and non-structural 2C protein were expressed and purified using a glutathione-S-transferase (GST) fusion protein expression system. We used a full-size CBV-4 cDNA as a template to amplify the genes by polymerase chain reaction (PCR). The genes were cloned into expression ve...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(97)00150-x
更新日期:1997-12-01 00:00:00
abstract::Laser microdissection combined with real-time RT-PCR presents an advanced tool to quantify particular RNA species in defined tissue areas. Dealing with infectious tissue samples increases the need to overcome the risk of infectivity and contamination during laser microdissection. Here, an useful method to control infe...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.03.024
更新日期:2006-08-01 00:00:00
abstract::TaqMan Mismatch Amplification Mutation Assay (TaqMAMA) is a highly sensitive allelic discrimination method. The mismatch amplification mutation assay (MAMA) is based on preferential amplification of mutant allele by the 'MAMA' primer, which is designed to have two mismatches with the wild-type allele and only one mism...
journal_title:Journal of virological methods
pub_type: 杂志文章,随机对照试验
doi:10.1016/j.jviromet.2008.07.020
更新日期:2008-11-01 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::Adenoviruses are highly efficient vectors for gene transfer into brain cells. Restricting transgene expression to specific cell types and maintaining long-term expression are major goals for adenoviral-mediated gene transfer in the central nervous system. Human adenovirus type 5 (Ad5) mediated transgene expression is ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2012.07.012
更新日期:2012-12-01 00:00:00
abstract::A comparison study was performed between the PLEX-ID and the CDC RT-PCR method for the detection and identification of Influenza A viruses using nasopharyngeal samples (N=75) collected between January and May 2011. Overall agreement was 89.3% (67/75 kappa=0.57 95% CI 0.3-0.89). Positive percent agreement was 92.3% (60...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.06.006
更新日期:2013-10-01 00:00:00
abstract::In this study, we aim to determine what effects prolonged storage and repeated freeze/thaw cycles have on the stability of influenza A(H1N1)pdm09 (influenza A/H1N1)RNA. Cloned influenza A/H1N1 RNA transcripts were serially diluted from 8.0-1.0 log10 copies/μl. RT-qPCR was used to measure RNA loss in transcripts stored...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2017.05.018
更新日期:2017-09-01 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 RT-PCR approach for the direct detection and typing of human enteroviruses in the environment is described in this study. A semi-nested RT-PCR using COnsensus-DEgenerated Hybrid Oligonucleotide Primers (CODEHOP) designed from the VP2 genome region has been developed for the direct typing of enteroviruses in clinical...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.07.018
更新日期:2014-10-01 00:00:00
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::The use of reporter constructs of HIV-1 molecular clones has been useful in studying the effects of infection on individual cells. A strategy of producing viral half-genome constructs containing reporter genes that can be used to produce recombinant complete infectious virions is described. This approach allows rapid ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(03)00110-1
更新日期:2003-06-30 00:00:00
abstract::An efficient method for construction of random epitope libraries using filamentous phage is described. Random DNA fragments generated by DNase I digestion were blunt ended by T4 DNA polymerase and ligated with a 12-mer linker, followed by PCR amplification using the same oligonucleotide linker as primers. The results ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(96)02057-5
更新日期:1996-07-01 00:00:00
abstract::Allele-specific PCR based on subtype consensus sequences is a powerful technique for detecting low frequency drug resistant mutants in HIV-1 infected patients. However, this approach can be limited by genetic variation in the region complementary to the primers, leading to variability in allele detection. The goals of...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2009.11.025
更新日期:2010-03-01 00:00:00
abstract::Bovine Papillomavirus type 2 (BPV-2) and chronic intoxication by bracken fern ingestion were associated with urinary bladder lesions and the clinical signs of enzootic haematuria in adult cattle. Clinically enzootic haematuria is characterized by intermittent haematuria followed by animal death. Enzootic haematuria ca...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.01.021
更新日期:2005-06-01 00:00:00
abstract::An enzyme avidin-biotin assay for the detection of anti-HBcAg antibody in human sera was developed. The assay uses genetically engineered HBcAg. HBcAg is immobilized on the surface of the wells of microtitre plates and the test serum sample, biotin-labelled HBcAg and streptavidin-labelled horseradish peroxidase are ad...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(89)90044-x
更新日期:1989-06-01 00:00:00
abstract::This study describes the development and evaluation of a cost effective test rationale for the detection of anti-HCV in dried blood spots. Samples were screened using an 'in house' IgG ELISA that incorporated the recombinant proteins c22-3, c200 and NS5. Confirmation of specific antibody to HCV was by a modification o...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(97)00127-4
更新日期:1997-11-01 00:00:00
abstract::A TaqMan real-time RT-PCR was developed to detect and quantify RNA-targets from the non-circulative, non-persistently transmitted Plum pox virus (PPV) in individual fresh or aphids captured previously and squashed on paper. Reliable quantitation ranged from 40 up to 4 x 10(8) copies of control transcripts. This techni...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.05.011
更新日期:2005-09-01 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::A protocol suitable for the detection of rabies virus and the related European bat lyssaviruses type 1 and 2 is described. In situ hybridisation, employing digoxigenin labelled riboprobes was used for the detection of lyssavirus RNA in mouse-infected brain tissue. The principal advantage of this technique, compared to...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2004.06.020
更新日期:2004-11-01 00:00:00
abstract::IgM antibody capture radioimmunoassay (MACRIA) and enzyme immunoassay (MACEIA) were compared with immunofluorescence (IF) for detecting specific IgM antibody in 99 sera from 76 infants with confirmed congenital rubella, and 61 sera from a comparative group of 59 infants who had miscellaneous abnormalities but in whom ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(82)90055-6
更新日期:1982-08-01 00:00:00
abstract::We have developed a sensitive method to detect and localize HPV-6 viral DNA, mRNA and protein in biopsy specimens of genital and respiratory tract lesions by using in situ hybridization and immunoperoxidase assays on sections of plastic-embedded tissue. This modified in situ hybridization technique, using ultrathin se...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(89)90098-0
更新日期:1989-07-01 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::Canine parvovirus (CPV) is an important disease causing gastroenteritis and/or haemorrhagic gastroenteritis in dogs. There are four antigenic types of CPV reported worldwide viz. CPV 2, CPV 2a, CPV 2b and CPV 2c. The diagnosis of CPV with the identification of the antigen type responsible remains problematic. In the p...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2016.02.013
更新日期:2016-07-01 00:00:00
abstract::Adeno-associated virus (AAV) is making its place in gene therapy applications; however, the industry is still facing obstacles in producing a large quantity of highly purified material for clinical studies. Insect cell technology can be used to produce AAV to meet the current demand. During the purification process it...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.09.011
更新日期:2007-01-01 00:00:00
abstract::The GPRIME (Group PRIMEr design) programs examine aligned sets of gene sequences to discover homologous regions to be targeted in diagnostic tests. The core program moves a 'window' over the aligned sequences and calculates, at each window position, a 'redundancy value', namely the number of sequences that would repre...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(98)00070-6
更新日期:1998-09-01 00:00:00
abstract::The plaque assay is essential for virion quantitation but the classic protocol requires considerable efforts. A simplified dengue 96-well plaque assay with automated quantitation program is an alternative to access the level of infectious virus. Dengue plaque assay was simplified using LLC/MK2 cells and virus mixing s...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2016.08.009
更新日期:2016-11-01 00:00:00
abstract::Among the enteric viruses implicated in waterborne outbreaks, human norovirus and hepatitis A virus (HAV) are a serious public health issue. Most foodborne viruses are difficult or currently unlikely to cultivate. Because of the lack of a cell culture method, real-time reverse transcriptase PCR is commonly used for th...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2020.113939
更新日期:2020-10-01 00:00:00
abstract::Six single-chain fragment variable (scFv) antibodies against infectious haematopoietic necrosis virus (IHNV) were selected from an antibody phage display library by phage display technology. The soluble scFv antibodies showed a molecular weight 32kDa by Western blot. Dot blot analysis revealed that the six scFv antibo...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.04.008
更新日期:2014-09-01 00:00:00