Abstract:
:The aim of this study was to develop a rapid, cost-saving triple reverse transcription polymerase chain reaction (triple RT-PCR) for subtyping H9N2 avian influenza viruses (AIVs). The three primer pairs for amplification of target sequences of nucleoprotein (NP), hemagglutinin (HA) and neuraminidase (NA) genes, respectively, were designed for subtyping the viruses in the triple RT-PCR. The sensitivity of triple RT-PCR was found to be 10(2) copies per reaction for each of NP, H9 and N2 gene. The specificity tests indicated that all of NP, HA and NA genes were positive for H9N2, only NP gene was positive for H5N1 and H1N1 AIVs, and the results were negative for the other avian viruses including Newcastle disease virus, infectious bronchitis virus, infectious bursal disease virus, duck hepatitis virus and avian encephalomyelitis virus. A total of 112 clinical samples were evaluated by the assay and the results showed that the sensitivity and specificity of triple RT-PCR were in accordance with the virus isolation. In conclusion, this method is rapid and cost-effective making it feasible and attractive for large-scale epidemiological investigation of H9N2 influenza virus.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Chen HT,Zhang J,Ma LN,Ma YP,Ding YZ,Wang M,Liu XT,Zhang YG,Liu YSdoi
10.1016/j.jviromet.2009.01.026subject
Has Abstractpub_date
2009-06-01 00:00:00pages
58-62issue
1-2eissn
0166-0934issn
1879-0984pii
S0166-0934(09)00019-6journal_volume
158pub_type
杂志文章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::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::The polymerase chain reaction (PCR) and restriction enzyme analysis were used to develop a rapid and simple procedure for identifying geographic subgroups of dengue virus within serotypes for epidemiologic investigations. The entire structural protein region of dengue viruses was amplified and the products were digest...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(94)90122-8
更新日期:1994-07-01 00:00:00
abstract::A newly developed real-time RT-polymerase chain reaction assay for quantitation of hepatitis C virus (HCV) RNA in human plasma and serum was applied. A pair of primers and a probe (molecular beacon) were designed that are specific for the recognition of a highly conservative 5'-non-coding region (5'-NCR) in HCV genome...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(02)00007-1
更新日期:2002-04-01 00:00:00
abstract::One of the major factors determining the incidence of Barley yellow dwarf virus (BYDV) on autumn-sown cereals is the viruliferous state of immigrant winged aphids. This variable is assessed routinely using the enzyme-linked immunosorbant assay (ELISA). However, the threshold for virus detection by ELISA can lead to fa...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(03)00097-1
更新日期:2003-06-09 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::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::Reverse genetics system is a powerful tool to study the function of a particular gene. The currently available reverse genetics system for Novirhabdovirus is based on vaccinia-driven T7 RNA polymerase expression. An improved system for the recovery of infectious hematopoietic necrosis virus (IHNV) was developed which ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2010.03.023
更新日期:2010-08-01 00:00:00
abstract::A method for duplex nested RT-PCR (nRT-PCR) with internal control (IC) for the detection of Nipah virus RNA is described. Incorporation of IC RNA distinguished false and true negative results. The extrinsic RNA was added directly to the PCR master mix and co-amplified with virus specific RNA in a duplex reaction to de...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2006.11.023
更新日期:2007-04-01 00:00:00
abstract::5-min boiling of 3-fold diluted serum leads to the quantitative release of immune-complexed HIV antigen (Ag) that can be readily measured. The method is capable of detecting as little as 4 to 5 pg/ml Ag p24 depending on the Ag test used. Of a panel of 50 anti-HIV-positive sera, 30 (60%) were Ag-positive after heat den...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(93)90080-b
更新日期:1993-07-01 00:00:00
abstract::The results of a comparison of three DNA-detection methods for human parvovirus B19 DNA are described. The sensitivity of detection of virus from hybridization assays using 32P-radiolabeled DNA and RNA probes was compared with a method for enzymatically amplifying specific target DNA sequences (polymerase chain reacti...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(89)90085-2
更新日期:1989-01-01 00:00:00
abstract::Lentiviral vectors (LVs) are promising delivery systems for gene therapy. To enhance the efficiency of target cell transduction by LVs, protocols often include the addition of culture additives. In this study, the cationic amphipathic peptide LAH4-L1 (KKALLAHALHLLALLALHLAHALKKA), a DNA transfection agent, was evaluate...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.02.005
更新日期:2013-05-01 00:00:00
abstract::In this inhibition immunoassay undiluted serum reacts in solution with crude cellular CMV antigen in wells of microtestplates coated with hyperimmune CMV-reactive monkey IgG. CMV antibodies in the serum under test block (completely or partial) the fixation of antigen to the capture layer. Unblocked antigenic activity ...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(87)90004-8
更新日期:1987-06-01 00:00:00
abstract::The coding sequence for a series of six histidines (his-tag) was inserted near the 5' terminus of the helper component (HC) coding region of tobacco etch potyvirus (TEV). Full length genomic clones containing the his-tag coding sequence were infectious and produced symptoms in tobacco (Nicotiana tabacuma) similar to t...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(98)00085-8
更新日期:1999-01-01 00:00:00
abstract::Peptides of HIV sequences are significant for antibody screening systems, and because of the limited number of amino acids they have to represent immunodominant regions of the virus proteins in order to maintain sensitivity. We detected, in a region of the outside loop of the transmembrane protein gp41 of the human im...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(89)90030-x
更新日期:1989-08-01 00:00:00
abstract::The sensitivity and specificity of four modes of two assays, immediate early nuclear antigen detection in cell culture (IENAD) at 24 and 48 h post-infection (p.i.) by immunofluorescence using a murine monoclonal antibody, and dot-blot DNA hybridisation with overnight or prolonged autoradiography using the 32P-labelled...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(87)90109-1
更新日期:1987-10-01 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::To develop reverse genetics system of RNA viruses, cloning of full-length viral genome is required which is often challenging due to many steps involved. In this study, we report cloning of full-length cDNA from an Indian field isolate (CSFV/IVRI/VB-131) of classical swine fever virus (CSFV) using in vitro overlap ext...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2015.10.006
更新日期:2015-12-15 00:00:00
abstract::Most transplant centers screen kidney transplant recipients for BK virus (BKV) infection using molecular techniques for the virus load determination. However, there is no consensus about the pre-analytical methods involved in the viral detection. In this study BK viral load was compared by the means of two urine treat...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2012.12.006
更新日期:2013-03-01 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::An indirect fluorescent antibody test (IFA) was evaluated using commercial mouse anti-measles monoclonal antibody and FITC-labeled goat anti-mouse immunoglobulin. For measles isolation, specimens were inoculated into Rhesus monkey kidney (RMK) cells and, when available, CV-1 cells. 381 specimens were tested by IFA and...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(91)90022-r
更新日期:1991-06-01 00:00:00
abstract::A genetically modified cell line (Vero-ICP10-EGFP) was constructed for detection of herpes simplex virus (HSV) by a simple, rapid and direct method. The cell line was developed by stable transfection of Vero cell with a plasmid encoding the green fluorescent protein (GFP) driven by the promoter of the HSV-2 ICP10 gene...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(00)00234-2
更新日期:2000-11-01 00:00:00
abstract::A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the detection of Cucurbit chlorotic yellows virus (CCYV). In this procedure, a set of four primers matching a total of six sequences in the coat protein gene region of CCYV was synthesized for the RT-LAMP assay using total...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.08.037
更新日期:2014-01-01 00:00:00
abstract::Congenital human cytomegalovirus infections are the major infectious cause of birth defects in the United States. How this virus crosses the placenta and causes fetal disease is poorly understood. Guinea pig cytomegalovirus (GPCMV) is a related virus that provides an important model for studying cytomegaloviral congen...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.01.031
更新日期:2008-05-01 00:00:00
abstract::A chimeric porcine circovirus type 1-2 (PCV1-2) infectious DNA clone has low transfection efficiency and exhibits low levels of proliferation. Electroporation and lipofection parameters were optimized for PK-15 and Dulac cells with the purpose of increasing the efficiency for rescuing infectious PCV1-2. Titers of PCV1...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2014.08.005
更新日期:2014-11-01 00:00:00
abstract::The performance of the new version of RT-PCR assay (Amplicor HIV-1 Monitor v1.5) was assessed. The quantification of non-B subtype HIV-1 plasma RNA (30A, 1C, 1D, 3E, 2F, 3G) obtained using Monitor v1.5 was compared to the former version of this assay (Monitor v1.0) and to the Quantiplex v2.0 bDNA assay. The new primer...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/s0166-0934(99)00057-9
更新日期:1999-08-01 00:00:00
abstract::Rice stripe virus (RSV), a planthopper-transmitted virus, was inoculated into rice protoplasts, and a one-step growth curve was determined. The amount of virus in the protoplasts decreased following the inoculation, and then increased after 8 h. The replication of RSV reached its peak 20 h after inoculation. RSV repli...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/0166-0934(96)02013-7
更新日期:1996-05-01 00:00:00
abstract::Mammalian orthoreoviruses (reoviruses) have served as highly useful models for studies of virus replication and pathogenesis. The development of a plasmid-based reverse genetics system represented a major breakthrough in reovirus research. The current reverse genetics systems for reoviruses rely on the expression of T...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2013.10.023
更新日期:2014-02-01 00:00:00
abstract::Hepatitis B-virucidal testing of biocides in quantitative suspension tests using duck hepatitis B virus (DHBV) requires primary duck embryonic hepatocytes for viral propagation. To improve the test system and availability of these cells, commercial culture plates with different growth surfaces were tested for cell cul...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2008.12.004
更新日期:2009-04-01 00:00:00
abstract::A nanoparticle-based immunoassay was developed for the rapid and sensitive detection of avian influenza virus (AIV). In this method, AIV-specific pentabody (pVHH3B) was conjugated to magnetic nanoparticles (MNPs) and used to capture AIV. Gold nanoparticles (GNPs), labelled with the anti-AIV mouse monoclonal antibody 3...
journal_title:Journal of virological methods
pub_type: 杂志文章
doi:10.1016/j.jviromet.2010.07.024
更新日期:2010-11-01 00:00:00