Generation of a human hepatoma cell line supporting efficient replication of a lamivudine resistant hepatitis B virus.


:Emergence of lamivudine (LAM) resistance causes treatment failure in patients with chronic hepatitis B and compromise the efficacy of subsequent salvage therapies with other nucleot(s)ide analogs (NAs). Establishment of cell-based assays supporting LAM-resistant hepatitis B virus (HBV) replication will not only provide tools for investigating the replication property, but also screening for antiviral agents efficiently inhibiting the replication of LAM-resistant HBV variants. Accordingly, a human hepatoma (HepG2)-derived cell line was established by stable transfection of a plasmid containing a 1.2 unit length of HBV genome harboring rtL180M and rtM204V mutations that confer LAM resistance. In addition to support efficient viral genome replication, the cell line also produces high levels of HBV virions and subviral particles. As expected, HBV DNA replication in this cell line is completely resistant to lamivudine, but sensitive to adefovir (ADV), entecavir (ETV) and tenofovir (TDF). The cell line is suitable for screening for antiviral agents that inhibit LAM-resistant HBV replication and inhibitors of HBsAg biosynthesis and secretion, which may reduce HBsAg antigenemia and ultimately help to restore host antiviral immune response against HBV and cure chronic HBV infection.


J Virol Methods


Zhang Y,Zhang Y,Kang Y,Wang J,Liu H,Zhu H,Qin Y,Mao R,Lin X,Lu M,Zhang J




Has Abstract


2014-06-01 00:00:00












  • Detection of the bovine herpesvirus-1 (BHV-1) genome by PCR.

    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


    authors: Vilcek S

    更新日期:1993-02-01 00:00:00

  • On the continuous culturing of B.95-8 cells in the presence of phosphonoacetic acid.

    abstract::Lymphoblastoid B.95-8 cells were cultured for four months and three weeks in the presence of increasing concentrations (50--200 microgram/ml) of phosphonoacetic acid (PAA). Several weeks after removal of the PAA, the cultures, in parallel with untreated B.95-8 cells, were tested for the presence of: 1) Epstein-Barr vi...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Margalith M,Manor D,Goldblum N

    更新日期:1980-01-01 00:00:00

  • Comparison of hemagglutination inhibition assay, an ELISA-based micro-neutralization assay and colorimetric microneutralization assay to detect antibody responses to vaccination against influenza A H1N1 2009 virus.

    abstract::The hemagglutination inhibition (HI) assay has been the main method used to investigate immune responses to vaccination against influenza H1N1 (2009) virus. However microneutralization tests (MNT) have been shown to be more sensitive and more specific. In this study, the three methods of choice: (i) the HI assay, (ii)...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Grund S,Adams O,Wählisch S,Schweiger B

    更新日期:2011-02-01 00:00:00

  • A new finding concerning adenoviral-mediated gene transfer: a high-level, cell-specific transgene expression in the neural stem cells of adult mice.

    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


    authors: Yan R,Zhang L,Zhang Q,Li J,Kang X,Wang H,Zhang J,Yang S,Yang X

    更新日期:2012-12-01 00:00:00

  • A novel next generation sequencing assay as an alternative to currently available methods for hepatitis C virus genotyping.

    abstract::Chronic HCV infection is one of the leading causes of liver-related death and in many countries it is a primary reason for having a liver transplant. HCV genotype identification has long been used in the clinical practice, since different genotypes have different response rates and required different doses and duratio...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Dirani G,Paesini E,Mascetra E,Farabegoli P,Dalmo B,Bartolini B,Garbuglia AR,Capobianchi MR,Sambri V

    更新日期:2018-01-01 00:00:00

  • Delipidation of a hepadnavirus: Viral inactivation and vaccine development.

    abstract::Many viruses including HIV, hepatitis C and hepatitis B, have an outer lipid envelope which maintains inserted viral peptides in the "correct" functional conformation and orientation. Disruption of the lipid envelope by most solvents destroys infectivity and often results in a loss of antigenicity. This communication ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Cham BE,Vickery K,Tohidi-Esfahani R,Cossart Y

    更新日期:2006-10-01 00:00:00

  • Development and validation of a novel SYBR Green real-time RT-PCR assay for the detection of classical swine fever virus evaluated on different real-time PCR platforms.

    abstract::Classical swine fever is a highly contagious viral disease that causes significant economic losses in pig production on a global scale. The rapid dissemination of the virus and the variability of the clinical signs merit the development of swift and accurate classical swine fever virus (CSFV) detection methods, which ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Pérez LJ,Díaz de Arce H,Tarradas J,Rosell R,Perera CL,Muñoz M,Frías MT,Nuñez JI,Ganges L

    更新日期:2011-06-01 00:00:00

  • Diagnosis of measles by fluorescent antibody and culture of nasopharyngeal secretions.

    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


    authors: Smaron MF,Saxon E,Wood L,McCarthy C,Morello JA

    更新日期:1991-06-01 00:00:00

  • Comparison of dot-blot DNA hybridisation and immediate early nuclear antigen production in cell culture for the rapid detection of human cytomegalovirus in urine.

    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


    authors: Morris DJ,Lomax J,Fox AJ,Corbitt G

    更新日期:1987-10-01 00:00:00

  • Immunogenicity and protective efficacy of a replication-defective infectious bronchitis virus vaccine using an adenovirus vector and administered in ovo.

    abstract::In ovo vaccination remains an attractive option for a cost effective, uniform and mass application of vaccines for commercial poultry. However, the vaccines which can be delivered safely by this method are limited and there is no currently licensed embryo-safe vaccine against infectious bronchitis virus (IBV). In this...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Zeshan B,Zhang L,Bai J,Wang X,Xu J,Jiang P

    更新日期:2010-06-01 00:00:00

  • Radioimmunoassay of adjuvant-associated porcine parvovirus using a monoclonal antibody in a nitrocellulose membrane system.

    abstract::A quantitative and simple indirect radioimmunoassay (IRIA) was developed for porcine parvovirus (PPV), employing a monoclonal antibody directed against PPV adsorbed to nitrocellulose membrane. The IRIA was equally sensitive to live or inactivated PPV. There was a linear relationship between membrane-bound radioactivit...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Katz JB,van Deusen RA

    更新日期:1985-12-01 00:00:00

  • A high-throughput assay for HIV-1 integrase 3'-processing activity using time-resolved fluorescence.

    abstract::HIV-1 integrase (HIV-1 IN), a well-validated antiviral drug target, catalyzes multistep reactions to incorporate viral DNA into the genome of the host cell; these include both a 3'-processing (3'P) reaction and a strand transfer reaction. These enzymatic activities can be measured in vitro with short DNA oligonucleoti...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Han YS,Quashie P,Mesplede T,Xu H,Mekhssian K,Fenwick C,Wainberg MA

    更新日期:2012-09-01 00:00:00

  • A new system to measure and compare hepatitis C virus replication capacity using full-length, replication competent viruses.

    abstract::Measuring the in vitro replication capacity of viruses is an important tool for assessing the effects of selective pressure of immune responses and drug therapy. Measuring hepatitis C virus (HCV) replication capacity utilizing primarily sub-genomic reporter constructs is limited. To overcome some of these limitations ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Lassmann B,Arumugaswami V,Chew KW,Lewis MJ

    更新日期:2013-12-01 00:00:00

  • A quantitative real-time reverse transcription PCR (qRT-PCR) assay to detect genome segment 9 of all 26 bluetongue virus serotypes.

    abstract::Bluetongue (BT) is an arboviral disease, which can often be fatal in naïve sheep and white tailed deer, but is usually less severe, or unapparent in other ruminants. Twenty-six bluetongue virus (BTV) serotypes have been recognised so far, two of which (BTV-25 and BTV-26) were recently identified by phylogenetic compar...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Maan NS,Maan S,Belaganahalli M,Pullinger G,Montes AJ,Gasparini MR,Guimera M,Nomikou K,Mertens PP

    更新日期:2015-03-01 00:00:00

  • Purification of the major envelop protein GP5 of porcine reproductive and respiratory syndrome virus (PRRSV) from native virions.

    abstract::Porcine reproductive and respiratory syndrome virus (PRRSV) is the cause of an economically important swine disease that has been devastating the global swine industry since the early 1990s. The current PRRSV vaccines are not very effective largely due to heterogeneic nature of the virus. The major envelope protein, G...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Matanin BM,Huang Y,Meng XJ,Zhang C

    更新日期:2008-01-01 00:00:00

  • Rescue of measles virus using a replication-deficient vaccinia-T7 vector.

    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


    authors: Schneider H,Spielhofer P,Kaelin K,Dötsch C,Radecke F,Sutter G,Billeter MA

    更新日期:1997-02-01 00:00:00

  • Analysis of varicella-zoster virus and herpes simplex virus in various clinical samples by the use of different PCR assays.

    abstract::Rapid and reliable detection of varicella-zoster virus (VZV) and herpes simplex virus type 1 (HSV-1) and -2 (HSV-2) is of clinical significance in immunocompromised patients and patients with infections of the central nervous system. This paper describes the detection of VZV and HSV using the commercially available Af...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Finnström N,Bergsten K,Ström H,Sundell T,Martin S,Wutzler P,Sauerbrei A

    更新日期:2009-09-01 00:00:00

  • A quantitative assay for measuring human foamy virus using an established indicator cell line.

    abstract::In order to improve the accuracy for detecting human foamy virus (HFV), an indicator cell line was established by co-transfecting baby hamster kidney-21 cells with two plasmids: one containing a G418 antibiotic resistance marker and the other including the luc gene which was placed downstream of the inducible HFV long...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Tai HY,Sun KH,Kung SH,Liu WT

    更新日期:2001-05-01 00:00:00

  • A rapid procedure for detecting noroviruses from cheese and fresh lettuce.

    abstract::Noroviruses (NoVs) are recognized as the most common agents of outbreaks of food-borne viral gastroenteritis and the efficiency of different methods for detection of NoVs from food matrices have been tested in several laboratories worldwide. The aim of this study was to develop a rapid and sensitive method for recover...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Fumian TM,Leite JP,Marin VA,Miagostovich MP

    更新日期:2009-01-01 00:00:00

  • Rapid synchronization of hepatitis C virus infection by magnetic adsorption.

    abstract::Hepatitis C virus (HCV) entry into target cells is thought to be a multistep process involving several cellular factors. However, their precise role during virus entry is unclear. Investigation of the mechanisms of HCV entry, such as the order of intervention by the cellular receptors, requires synchronizing infection...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Vieyres G,Angus AG,Haberstroh A,Baumert TF,Dubuisson J,Patel AH

    更新日期:2009-04-01 00:00:00

  • Development of a dot-immunobinding assay for detection of citrus tristeza virus.

    abstract::The dot-immunobinding assay (DIBA) was adapted for detection of citrus tristeza virus (CTV) and compared with DAS-ELISA and DAS-indirect ELISA. DIBA was easy to perform and as sensitive as either ELISA procedure for CTV diagnosis. The entire test could be performed in 2-3 h using polyclonal antibodies, with minimal la...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Rocha-Peña MA,Lee RF,Niblett CL

    更新日期:1991-10-01 00:00:00

  • Determination of specific cytomegalovirus IgM antibodies using infected air dried cells and isolated nuclei by immunoperoxidase assay.

    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


    authors: Singer Y,Kimmel N,Sarov I

    更新日期:1985-05-01 00:00:00

  • A solid phase reverse transcriptase micro-assay for the detection of human immunodeficiency virus and other retroviruses in cell culture supernatants.

    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


    authors: Somogyi PA,Gyuris A,Földes I

    更新日期:1990-03-01 00:00:00

  • Analysis of the expression profiles of Marek's disease virus-encoded microRNAs by real-time quantitative PCR.

    abstract::MicroRNAs (miRNAs) are a large class of endogenous approximately 22-nucleotide long non-coding RNAs involved in post-transcriptional silencing of gene expression by translational repression or direct cleavage of the target mRNAs. Several hundreds of miRNAs have now been identified in a wide range of organisms includin...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Xu H,Yao Y,Zhao Y,Smith LP,Baigent SJ,Nair V

    更新日期:2008-05-01 00:00:00

  • An international collaborative study to assess a set of reference reagents for HIV-1 PCR.

    abstract::An international collaborative study was performed to evaluate a set of PCR reference reagents for HIV diagnosis. Twenty-six laboratories from 9 countries analysed a proficiency panel of 10 coded DNA samples using the PCR reference reagents and protocols. For comparison, these coded samples were then assessed using a ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Bootman JS,Kitchin PA

    更新日期:1992-04-01 00:00:00

  • Chicken single-chain variable fragments against the SARS-CoV spike protein.

    abstract::The major concern for severe acute respiratory syndrome (SARS), caused by the SARS-associated coronavirus (SARS-CoV), is the lack of diagnostic and therapeutic agents. Using a phage display technology in a chicken system, high-affinity monoclonal antibody fragments against the SARS-CoV spike protein were characterized...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Lee YC,Leu SJ,Hu CJ,Shih NY,Huang IJ,Wu HH,Hsieh WS,Chiang BL,Chiu WT,Yang YY

    更新日期:2007-12-01 00:00:00

  • Real-time PCR quantitation of hepatitis B virus total DNA and covalently closed circular DNA in peripheral blood mononuclear cells from hepatitis B virus-infected patients.

    abstract::It remains unclear whether hepatitis B virus (HBV) replicates in extrahepatic tissues, and particularly in peripheral blood mononuclear cells (PBMCs), which may serve as a reservoir for the maintenance of infection. A real-time PCR assay for the quantitation of total and covalently closed circular (ccc) HBV DNA in ser...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Mazet-Wagner AA,Baclet MC,Loustaud-Ratti V,Denis F,Alain S

    更新日期:2006-12-01 00:00:00

  • Preservation of RNA and destruction of infectivity in microdissected brain tissues of Lewis rats infected with the Borna disease virus.

    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


    authors: Porombka D,Herzog S,Baumgärtner W,Herden C

    更新日期:2006-08-01 00:00:00

  • Comparison of specific and random priming in the reverse transcriptase polymerase chain reaction for genotyping group A rotaviruses.

    abstract::This study describes an approach to the molecular typing of rotaviruses which requires only a single RNA extraction and reverse transcription (RT) reaction using random primers. Random-primed RT provides complementary DNA (cDNA) which can be used not only for G- and P-typing polymerase chain reactions (PCR), but also ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Iturriza-Gomara M,Green J,Brown DW,Desselberger U,Gray JJ

    更新日期:1999-03-01 00:00:00

  • A non-radioisotopic reverse transcriptase assay using biotin-11-deoxyuridinetriphosphate on primer-immobilized microtiter plates.

    abstract::We developed a non-radioisotopic (non-RI) reverse transcriptase assay (RTA). The reverse transcriptase (RT) incorporates biotin-11-deoxyuridine-triphosphate (bio-dUTP) using a poly(rA) template hybridized with oligo(dT) primer that is immobilized on the surface of a 96-well microtiter plate. This assay is thus semi-au...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Urabe T,Sano K,Tanno M,Mizoguchi J,Otani M,Lee MH,Takasaki T,Kusakabe H,Imagawa DT,Nakai M

    更新日期:1992-11-01 00:00:00