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 sections and strand-specific 3H-labelled riboprobes, offers the advantages of superior morphological preservation and detection of viral genomes at low copy number with good resolution. This modified immunocytochemistry provides better sensitivity when compared to previous methods using paraffin-embedded materials. In respiratory tract lesions, immunoperoxidase assay detected only a few capsid antigen positive cells, while in the genital tract lesions, there were more capsid antigen positive cells. Southern transfer analyses and in situ hybridizations demonstrated the presence of more viral nucleic acids in genital tract papillomata than respiratory tract papillomata. Epithelial cells throughout the papillomata were infected by HPV-6 as evidenced by positive hybridization, with more viral DNA present in superficial cells. Our results suggest that genital tract epithelium is more permissive for HPV-6 replication than respiratory tract epithelium. Using stand-specific probes synthesized from subgenomic fragments of the HPV-6 genome in conjunction with nuclease digestions, we were able to demonstrate that HPV-6 transcripts specific to open reading frames (ORFs) E6, E7, E1, L1, and L2 occur in maturing superficial cells. In contrast, transcripts specific to ORFs E1, E2, E4, E5a, and E5b could be detected throughout the whole of the epithelium with more signals noted at the basal cell areas. In addition, the distribution of HPV-6 nucleic acids and protein in a carcinoma in situ of the larynx was analyzed. In comparison to benign respiratory tract papillomata, more viral DNA was found in the malignant lesion, but the pattern and distribution of transcription and capsid antigen was similar.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Wu TC,Mounts Pdoi
10.1016/0166-0934(89)90098-0subject
Has Abstractpub_date
1989-07-01 00:00:00pages
31-47issue
1eissn
0166-0934issn
1879-0984pii
0166-0934(89)90098-0journal_volume
25pub_type
杂志文章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 real-time reverse transcription-polymerase chain reaction (qRT-PCR) assay was developed for the fast and highly sensitive detection of the sacbrood virus (SBV) genome and applied to honeybee samples. Using plasmid DNA containing a partial SBV genome and diluted serially, as few as 1×10(2)copies/μl (correlation co-ef...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2011.10.014
更新日期:2012-01-01 00:00:00
abstract::Dengue surveillance relies on reverse transcription-polymerase chain reaction (RT-PCR), for confirmation of dengue virus (DENV) serotypes. We compared efficacies of published and modified primer sets targeting envelope (Env) and capsid-premembrane (C-prM) genes for detection of circulating DENV serotypes in southern I...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2017.02.014
更新日期:2017-06-01 00:00:00
abstract::Plum pox virus (PPV) causes the most destructive viral disease known as plum pox or Sharka disease in stone fruit trees. As an important regulated pathogen, detection of PPV is thus of critical importance to quarantine and eradication of the spreading disease. In this study, the innovative development of two AmplifyRP...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.06.026
更新日期:2014-10-01 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::The in vitro syncytium induction capacity of human immunodeficiency virus type 1 (HIV1) isolates is an important marker in the progression of the disease. Two methods have been widely used to determine the biological phenotype of HIV1. These two methods, the direct MT-2 assay and the supernatant assay, were compared f...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(96)02152-0
更新日期:1997-03-01 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::Cell culture based assays used to detect waterborne viruses typically call for incubating the sample for at least two weeks in order to ensure that all the culturable virus present is detected. Historically, this estimate was based, at least in part, on the length of time used for detecting poliovirus. In this study, ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2016.09.002
更新日期:2016-11-01 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::We assessed the performance of Abbott RealTime CMV assay (ARC) compared to Roche Cobas Amplicor CMV Monitor Test (RCM) for quantification of CMV in plasma of transplant patients. Commercial panels were used to test linearity, precision and interference and 83 clinical samples were used for the accuracy and precision a...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2015.08.010
更新日期:2015-12-01 00:00:00
abstract::This paper describes the cloning, sequencing and bacterial expression of the N protein of the Rift Valley fever virus (RVFV) ZIM688/78 isolate and its evaluation in indirect ELISAs (I-ELISA) for the detection of IgM and IgG antibodies in human and sheep sera. Sera used for the evaluation were from 106 laboratory worke...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.11.005
更新日期:2007-03-01 00:00:00
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::In the context of eradication of poliomyelitis the World Health Organization stimulates the development of inactivated polio vaccines based on attenuated virus strains. In addition to vaccine development, tests have to be designed to assess the vaccine quality. An important test is the identification test for poliovir...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.01.026
更新日期:2013-04-01 00:00:00
abstract:BACKGROUND:Diagnostic real time reverse transcription PCR (rRT-PCR) is usually done using nucleic acid (NA) purified from the sample. In the SARS-CoV-2 pandemic reagents and utensils for NA purification has been in short supply. This has generated interest in methods that eliminate the need for NA purification. OBJECT...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2021.114062
更新日期:2021-01-08 00:00:00
abstract::An indirect enzyme-linked immunosorbent assay termed rnPRRS ELISA using baculovirus-expressed and affinity-purified viral nucleocapsid protein (rNC) antigen was developed for detecting antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) in swine sera. Sera (1395) originating from different E...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(97)02186-1
更新日期:1997-05-01 00:00:00
abstract::A lateral flow device (LFD) for the detection of swine vesicular disease (SVD) virus (SVDV) and differential diagnosis from foot-and-mouth disease (FMD) was developed using a monoclonal antibody (Mab C70). The performance of the LFD was evaluated in the laboratory on suspensions of vesicular epithelia and cell culture...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2009.09.023
更新日期:2010-02-01 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::A simple sequencing-based assay is described for genotyping of hepatitis C virus (HCV). RT-PCR was employed to amplify a 237-nucleotide-long fragment from the 5' untranslated region (UTR) of the genome using one biotinylated and one normal primer. Subsequent to capture of the PCR products on streptavidin-coated beads,...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(03)00068-5
更新日期:2003-05-01 00:00:00
abstract::SIV infection in macaques has become an important animal model for HIV-1 infection in humans. An antibody assay was therefore developed and compared to a commercially available antigen assay with respect to their usefulness to monitor the course of simian immunodeficiency virus (SIV) infection in cynomolgus monkeys. A...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(92)90019-a
更新日期:1992-04-01 00:00:00
abstract::Two genotype-specific fluorogenic RT-PCR assays were developed for the detection and quantitation of canine coronavirus (CCoV) type I and type II RNA in the faeces of dogs with diarrhoea. Both the fluorogenic assays showed high specificity, sensitivity and reproducibility, allowing a precise quantitation of CCoV type ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.06.005
更新日期:2005-12-01 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::The performance of the rapid confirmatory HIV 1/2 assay Geenius was compared with the conventional HIV 1/2 line immunoblot (INNO-LIA HIV I/II Score). One hundred HIV 1/2 confirmed positive samples from donors and patients and 136 negative screening samples from blood donors were evaluated with both assays. A 20 member...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.05.010
更新日期:2014-09-01 00:00:00
abstract::Clones isolated from a genomic library of the baculovirus PmSNPV (= MBV) were used to prepare DNA probes for detection of PmSNPV in shrimp tissue. Davidson's AFA fixed shrimp tissues were probed using DNA that had been labeled with the hapten, digoxigenin, by an in situ hybridization assay. The DNA probes readily dist...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(94)90043-4
更新日期:1994-09-01 00:00:00
abstract::In many mammals, viruses cause hepatitis. Despite many efforts a specific virus responsible for canine idiopathic hepatitis has not been identified. The discovery of a viral etiology in canine hepatitis will promote the development of specific drugs and vaccines for the treatment of idiopathic hepatitis in dogs. The o...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2012.05.019
更新日期:2012-10-01 00:00:00
abstract::In order to identify new potential antiviral drugs, small amounts of extracts or compounds have to be examined for cytotoxicity and antiviral activity in primary screening using a rapid, easy, inexpensive, and highly standardised test system. In this study, high-throughput cytopathic effect (CPE) inhibitory assays wer...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(01)00305-6
更新日期:2001-06-01 00:00:00
abstract::HCV-RNA was examined in serum and liver tissue obtained from 8 hepatitis B surface antigen (HBsAg) negative patients with liver nodules ranging in size from 2 to 11 cm. Histological examination of ultrasound-guided fine needle biopsies revealed the presence of hepatocellular carcinoma (HCC) in six patients (5 of whom ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(94)90112-0
更新日期:1994-07-01 00:00:00
abstract::Bovine herpesvirus 4 (BoHV-4) is a gamma herpesvirus with no clear disease association. Previous studies have demonstrated that macrophages can harbour persistent BoHV-4. Since mesenchymal stem cells in bone marrow regulate the differentiation and proliferation of adjacent haematopoietic precursors, such as macrophage...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2005.02.019
更新日期:2005-08-01 00:00:00
abstract::Hexon is the major structural protein of adenovirus, and has significance in studies of virus structure and function, vaccine development, and immunodiagnosis. We describe a simple, single-step, anion-exchange high performance liquid chromatography (HPLC) method for the high yield purification of hexon. Purity of the ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(87)90131-5
更新日期:1987-09-01 00:00:00
abstract::A simple method for evaluating the virucidal activity of water-soluble antiseptic and disinfectant products using poliovirus type 1 (Sabin strain) is described. Using a commercial concentrator, kinetic studies of four products were carried out. It was shown that 2% glutaraldehyde and 5% povidone iodine are rapidly vir...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(84)90052-1
更新日期:1984-12-01 00:00:00
abstract::A system which allows the reconstitution of measles virus (MV) from cloned cDNA is described. The severely host cell restricted vaccinia vector MVA-T7 expressing bacteriophage T7 RNA polymerase was used to generate full-length antigenomic MV RNA and simultaneously the mRNAs encoding the viral N, P and L proteins in or...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(96)02137-4
更新日期:1997-02-01 00:00:00