Abstract：
:To facilitate efficient and accurate detection of potato-infecting carlaviruses, degenerated universal primers were designed based on conserved amino acid and nucleotide sequences. Two sense primers, Car-F1 and Car-F2, were based on the amino acid sequences "SNNMA" and "GLGVPTE", respectively, in the coat protein. The reverse primer, Car-R, which was located at the border of the nucleic acid binding protein gene and the 3' untranslated region, and dT-B, which was derived from the oligo-dT targeting the poly(A) tail, were selected. Successful application of fragments within the predicted size range of carlaviruses was obtained using Car-F1 paired with either Car-R or dT-B from tested carlaviruses (Potato virus S, M and latent) by RT-PCR. The Car-F2 failed to yield clear-cut fragments within the predicted size range when paired with either Car-R or dT-B in RT-PCR. However, a less degenerated version of the primer, Car-F2b, resulted in amplicons within the predicted size range when paired with either Car-R or dT-B. Sequencing of the tentative carlavirus-fragments resulting from Car-F1/Car-R and Car-F2b/dT-B proved their carlavirus-origin, thus indicating the high specificity of these primers. The sensitivity of Car-F1/Car-R or Car-F2b/Car-R mediated RT-PCR for the detection of carlavirus-infected potato tubers were assessed using composite samples containing one carlavirus-infected-potato-tuber RNA sample with up to 49 virus-free-potato-tuber RNA samples under the optimal annealing temperature. The target carlaviruses were detected readily from all composites, demonstrating a high sensitivity. The method was further evaluated using presumed virus-free or carlavirus-infected potatoes of several cultivars, and reliable results were obtained.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Nie X,Bai Y,Molen TA,Desjardins DCdoi
10.1016/j.jviromet.2008.02.004subject
Has Abstractpub_date
2008-05-01 00:00:00pages
209-16issue
2eissn
0166-0934issn
1879-0984pii
S0166-0934(08)00066-9journal_volume
149pub_type
杂志文章abstract：:In preparation for field trials, a nuclear polyhedrosis virus (NPV) of the pine beauty moth, Panolis flammea, was mass produced in vivo in an alternative (heterologous) host, the cabbage moth, Mamestra brassicae. Simultaneously, Mamestra NPV was also produced in M. brassicae. This homologous NPV/host system was a cons...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(88)90019-5
更新日期：1988-03-01 00:00:00
abstract：:Group A rotaviruses (RVA) are important infectious agents associated with diarrhea in the young of several animal species including foals. Currently, a variety of diagnosis methods are commercially available, like ELISA, latex agglutination and immunochromatographic assays. These commercial tests are mainly designed f...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2015.05.002
更新日期：2015-09-15 00:00:00
abstract：:Sample preparation is an important step in the detection of viral DNA by the polymerase chain reaction (PCR) technique. The method used should achieve release of cellular DNA with the minimum of manipulation steps so as to reduce the possibility of contamination. The present report demonstrates that either microwaving...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90009-g
更新日期：1993-09-01 00:00:00
abstract：:Simian betaretroviruses (formerly Type D retroviruses; SRV) are a group of closely related retroviruses for which the natural host species are Asian monkeys of the genus Macaca. Five serotypes have been identified by classical neutralization assays and three additional untyped variants have been reported (SRV(Tsukuba)...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.07.030
更新日期：2009-12-01 00:00:00
abstract：:The availability of whole genome sequencing has contributed to many aspects of dengue research, and its use in dengue virus (DENV) surveillance for early epidemic warning has been proposed. Methods to sequence the genomes of individual dengue serotypes have been described previously, but no single method is known to b...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.06.013
更新日期：2010-10-01 00:00:00
abstract：:Rubella virosomes were prepared from performed liposomes and detergent solubilized viral hemagglutinin. The liposomes were made from lecithin/dicetyl phosphate (3.5 : 1) films resuspended in NTE buffer and sonicated. Viral hemagglutinin was prepared from purified virus after solubilization with Triton X-100 and centri...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(81)90069-0
更新日期：1981-11-01 00:00:00
abstract：:Numerous strategies have been employed in an attempt to improve the immunogenicity and efficacy of nucleic acid vaccines. In the present study, the immunogenicity in the induction of humoral and cellular immune responses to HIV-1 DNA vaccine expressing a chimeric gene of gag and gp120 and the adjuvant effect of IFN-al...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2007.07.016
更新日期：2007-12-01 00:00:00
abstract：:The study of replication of viruses that require high bio-secure facilities can be accomplished with less stringent containment using non-infectious 'replicon' systems. The FMDV replicon system (pT7rep) reported by Mclnerney et al. (2000) was modified by the replacement of sequences encoding chloramphenicol acetyl-tra...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2014.08.020
更新日期：2014-12-01 00:00:00
abstract：:A simple immunoperoxidase assay (IPA), adapted for detection of serum IgM antibodies to cytomegalovirus (CMV) is described. The antigen consisted of CMV infected human embryonic fibroblasts or isolated nuclei. The sera were absorbed with aggregated gamma-globulins prior to testing. Rabbit anti-human IgM peroxidase con...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(85)90122-3
更新日期：1985-05-01 00:00:00
abstract：:A liquid phase blocking sandwich ELISA has been compared with virus neutralisation for testing pig sera for antibodies against swine vesicular disease (SVD) virus. Highest infectivity titre of SVD virus was obtained using a multiplicity of infection of 30 pfu/cell and harvesting after 21 h. Titres obtained for 300 cli...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(89)90101-8
更新日期：1989-07-01 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：:Paramyxoviruses, including measles virus (MV), human metapneumovirus (HMPV), human respiratory syncytial virus (HRSV) and canine distemper virus (CDV), are transmitted via the respiratory route. Despite their close phylogenetic relationship, the pathogenesis of these viruses is very different. To study viral tropism a...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2013.06.016
更新日期：2013-10-01 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：:Type-specific 30'mer-36'mer oligonucleotide probes complementary to mRNA transcribed from the E6 and E7 open reading frames of HPV 6b/11, 16, 18 and 33 were designed using the published nucleotide sequences. As oligonucleotides are easily and relatively cheaply synthesized in large amounts and are free of vector DNA, ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(93)90018-m
更新日期：1993-03-01 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：:Following the introduction of African swine fever virus (ASFV) into Europe in 2007, ASFV infection has spread continuously over the past years and it became a high level disease threat in Europe and also Asia. Examination of suspect clinical cases for ASF with rapid and sensitive laboratory methods can substantially c...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2020.113886
更新日期：2020-05-23 00:00:00
abstract：:Codon-optimization refers to the alteration of gene sequences, to make codon usage match the available tRNA pool within the cell/species of interest. Codon-optimization has emerged as a powerful tool to increase protein expression by genes from small RNA and DNA viruses, which commonly contain overlapping reading fram...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(03)00173-3
更新日期：2003-08-01 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：:The role of environmental reservoirs in avian influenza virus (AIV) transmission has been investigated during AIV-associated outbreaks. To date, no method has been defined for detection of AIV from mud samples. A procedure using elution and polyethylene glycol (PEG) concentration steps was designed to detect AIV by RT...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2011.10.013
更新日期：2012-01-01 00:00:00
abstract：:Porcine reproductive and respiratory syndrome virus (PRRSV) has a marked tropism for cells of the monocyte-macrophage lineage and accordingly, replicates in fully differentiated alveolar macrophages in the natural host. Despite the identification of several putative receptors for PRRSV on porcine alveolar macrophages ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2009.11.003
更新日期：2010-02-01 00:00:00
abstract：:A method for detecting the emergence of potential pandemic-causing influenza strains has been developed. The system first uses real-time RT-PCR to detect H5, the highly pathogenic avian influenza subtype most likely to cause a pandemic. Pyrosequencing is then employed to scan for codon changes encoding amino acids kno...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2008.09.013
更新日期：2008-12-01 00:00:00
abstract：:Japanese encephalitis (JE) is caused by the Japanese encephalitis virus (JEV). It is a major public health problem in Asia. JEV infects swine which results in fatal encephalitis, abortion and stillbirth in pregnant sow, and hypospermia in boars. Swine is a viral amplifier, and thus plays a critical role in JEV transmi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.04.015
更新日期：2010-09-01 00:00:00
abstract：:Systematic solid-phase synthesis of all possible overlapping nonapeptides of the 1381 amino acid sequence of the Epstein-Barr virus major capsid protein (EBV-MCP) was used to identify the position of linear antigen epitopes on this protein as recognised by human polyclonal antisera. Antisera were selected for reactivi...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(88)90061-4
更新日期：1988-09-01 00:00:00
abstract：:The humoral immune response of sheep infected with bluetongue virus serotypes 3, 9 and 16 was monitored by plaque inhibition (PI), blocking ELISA (BELISA) and indirect ELISA over a period of 63 days post-infection. Results indicated that testing of a single plasma or serum sample by both a BELISA and an indirect ELISA...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/0166-0934(94)90088-4
更新日期：1994-06-01 00:00:00
abstract：:In the last decades, molecular techniques have gradually been adopted for the rapid confirmation of results obtained through gold standard methods. However, international organisations discourage their use in routine laboratory investigations for rabies post-mortem diagnosis, as they may lead to false positive results...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2016.03.017
更新日期：2016-08-01 00:00:00
abstract：:Louping ill-like virus (LI) has been recently detected in two different locations in the north of Spain and separated by only around 400 km. Using molecular approaches, the viruses causing both outbreaks have been shown to be different to LI virus, but also different to each other. They have been called SSEV (Spanish ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2018.02.017
更新日期：2018-05-01 00:00:00
abstract：:Laboratory analysis of blood specimens is an increasingly important tool for rapid diagnosis and control of therapy. So, miniaturization of test systems is needed, but reduced specimens might impair test quality. For rapid detection and quantitation of HBV DNA, the COBAS(®) AmpliPrep/COBAS(®) TaqMan(®) HBV test has pr...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2010.07.025
更新日期：2010-11-01 00:00:00
abstract：:In situ hybridization detection of viral RNAs in formaldehyde-fixed tissue specimens is used frequently to characterize the extent of viral replication within host tissues. The ability to determine the level of expression of viral RNAs in situ is dependent upon many factors including the extent of cross-linking during...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(01)00378-0
更新日期：2002-01-01 00:00:00
abstract：:Nucleic acid amplification techniques have become the mainstay for ultimate sensitivity for detecting low levels of virus, including human immunodeficiency virus (HIV). As a sophisticated technology with relative expensive reagents and instrumentation, adoption of nucleic acid testing (NAT) can be cost inhibited in se...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/j.jviromet.2012.08.017
更新日期：2013-03-01 00:00:00
abstract：:A recombinant retroviral system was used for the analysis of early HIV breakthrough infection in the presence of antiviral drugs. The use of replication-defective HIV allowed a quantitative analysis of a single cycle of infection. This report characterizes this recombinant HIV system and demonstrates it's validity in ...
journal_title：Journal of virological methods
pub_type： 杂志文章
doi：10.1016/s0166-0934(97)00212-7
更新日期：1998-04-01 00:00:00
