Analysis of unknown DNA sequences by polymerase chain reaction (PCR) using a single specific primer and a standardized adaptor.


:A new procedure for the PCR amplification of unknown DNA sequences adjacent to a known sequence is described. The required but not readily available second primer sequence in the unknown DNA sequence is obtained by creating an overhanging restriction site in the unknown sequence to which a double-stranded oligonucleotide adaptor of known sequence is ligated.


J Virol Methods


Collasius M,Puchta H,Schlenker S,Valet G




Has Abstract


1991-04-01 00:00:00














  • Screening of herpes simplex virus type 1 isolates for acyclovir resistance using DiviTum® assay.

    abstract::Rapid alternative methods are required to evaluate easily acyclovir (ACV) sensitivity of clinical herpes simplex virus (HSV) isolates. The objective of this study was to screen 54 ACV-sensitive and 41 ACV-resistant clinical HSV-1 isolates, well characterized by phenotypic and genotypic methods, for the phosphorylation...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Sauerbrei A,Vödisch S,Bohn K,Schacke M,Gronowitz S

    更新日期:2013-03-01 00:00:00

  • Real-time polymerase chain reaction assay based on high-resolution melting analysis for the determination of the rs12979860 polymorphism involved in hepatitis C treatment response.

    abstract::Treatment of chronic hepatitis C is associated with varying success rates, substantial medical costs and serious side effects. Several host polymorphisms have been identified near the IL28B gene, of which the homozygous rs12979860 CC was found to be associated with significantly favourable treatment outcome. To determ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Kovanda A,Poljak M

    更新日期:2011-07-01 00:00:00

  • Detection of group and interspecies reactivities of mammalian C-type virus p30 proteins by an enzyme-linked immunosorbent assay (ELISA): enhancement of interspecies reactivity by denaturation.

    abstract::The applicability of the enzyme immunoassay technique to the detection of group and interspecies determinants of C-type retroviral proteins was tested. For this purpose four groups of C-type retroviruses (MuLV, FeLV, SiSV/GaLV, and BaEV/RD114) were assayed for group-specific and interspecies reactivities of their p30 ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Schetters H,Hehlmann R,Erfle V,Saxinger C

    更新日期:1982-11-01 00:00:00

  • Molecular cloning of Indian tomato leaf curl virus genome following a simple method of concentrating the supercoiled replicative form of viral DNA.

    abstract::DNA-A and DNA-B components of the genome of a whitefly transmitted virus causing yellowing and leaf curl in tomato (ITLCV) were cloned following a simple procedure for isolation of the double stranded replicative form of viral DNA from infected tomato plants. The method is based on extraction of total DNA from infecte...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Srivastava KM,Hallan V,Raizada RK,Chandra G,Singh BP,Sane PV

    更新日期:1995-02-01 00:00:00

  • Development of a universal RT-PCR for amplifying and sequencing the leader and capsid-coding region of foot-and-mouth disease virus.

    abstract::Foot-and-mouth disease (FMD) is a highly infectious viral disease of cloven-hoofed animals with debilitating and devastating consequences for livestock industries throughout the world. Key antigenic determinants of the causative agent, FMD virus (FMDV), reside within the surface-exposed proteins of the viral capsid. T...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Xu L,Hurtle W,Rowland JM,Casteran KA,Bucko SM,Grau FR,Valdazo-González B,Knowles NJ,King DP,Beckham TR,McIntosh MT

    更新日期:2013-04-01 00:00:00

  • Sap-direct RT-PCR for the rapid detection of coleus blumei viroids of the genus Coleviroid from natural host plants.

    abstract::A simple and fast sap-direct RT-PCR (reverse transcription-polymerase chain reaction) for the rapid detection of 3 viroids of the genus Coleviroid is presented. The templates for cDNA synthesis were obtained directly from the sap of coleus using a pipettor, a common tool in molecular biology laboratories, and 3 coleus...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Jiang D,Wu Z,Xie L,Sano T,Li S

    更新日期:2011-06-01 00:00:00

  • Humoral response and genetic stability of recombinant BCG expressing hepatitis B surface antigens.

    abstract::The Bacille Calmette Guerin (BCG), long valued for its role as a live vaccine for the prevention of tuberculosis, is being used as a recombinant delivery vehicle for foreign antigens, principally, for inducing long-lived specific humoral and cellular immunity. Hepatitis B and its sequelae are major public health probl...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Rezende CA,De Moraes MT,De Souza Matos DC,McIntoch D,Armoa GR

    更新日期:2005-04-01 00:00:00

  • Sensitive detection of Tomato ringspot virus by real-time TaqMan RT-PCR targeting the highly conserved 3'-UTR region.

    abstract::A real-time TaqMan RT-PCR assay was developed for the rapid and sensitive detection of Tomato ringspot virus (ToRSV), an important plant virus which infects a wide range of fruit and ornamental crops. Primers and a probe were designed based on the highly conserved 3'-untranslated region (UTR) sequences of ToRSV, to am...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Tang J,Khan S,Delmiglio C,Ward LI

    更新日期:2014-06-01 00:00:00

  • Three unique Sendai virus antigenic peptides screened from nucleocapsid protein by overlapping peptide array.

    abstract::Sendai virus (SeV) is strictly monitored in laboratory rodents. Currently, complete virions have been used as antigens in SeV serological tests. However, the complexity of SeV virion antigen limits the accuracy of the diagnostic method. In the current study, complete SeV virion antigen was separated on SDS-PAGE and an...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Xiang Z,Tong W,Li Y,Qin C,Wei Q

    更新日期:2013-11-01 00:00:00

  • Simultaneous detection of five different DNA targets by real-time Taqman PCR using the Roche LightCycler480: Application in viral molecular diagnostics.

    abstract::One of the most interesting aspects of real-time PCR based on the detection of fluorophoric labeled oligonucleotides is the possibility of being able to detect conveniently multiple targets in the same PCR reaction. Recently, Roche Diagnostics launched a real-time PCR platform, the LightCycler480 (LC480), which should...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Molenkamp R,van der Ham A,Schinkel J,Beld M

    更新日期:2007-05-01 00:00:00

  • Development and application of a universal Taqman real-time PCR for quantitation of duck hepatitis B virus DNA.

    abstract::To develop a quantitative assay for universal detection of duck hepatitis B virus (DHBV) DNA, a Taqman real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) assay was developed using primers and probes based on genomic sequences located at nucleotide 241-414 of the DHBV Core region which possesses the ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Wang Y,Li Y,Yang C,Hui L,Han Q,Ma L,Wang Q,Yang G,Liu Z

    更新日期:2013-07-01 00:00:00

  • A procedure for the isolation of specific point mutants of influenza virus.

    abstract::Procedures were developed for the screening and isolation of RNA viruses that vary from the consensus population by a single point mutation and are present in low abundance. We tested these procedures using a mixture of the vaccine donor line cold-adapted (ca) B/Ann Arbor/1/66 and its wild type (wt) progenitor at a ra...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Mitchell BM,DeBorde DC

    更新日期:1993-05-01 00:00:00

  • A portable detection assay for Apple stem pitting virus using reverse transcription-recombinase polymerase amplification.

    abstract::A molecular diagnostic assay for the rapid, sensitive and specific detection of Apple stem pitting virus (ASPV) in infected samples, utilizing reverse transcription-recombinase polymerase amplification (RT-RPA) at an isothermal constant temperature of 42 °C and the designed target-specific primers, was developed. The ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Kim NY,Lee HJ,Jeong RD

    更新日期:2019-12-01 00:00:00

  • Immunogenicity and virulence of attenuated vaccinia virus Tian Tan encoding HIV-1 muti-epitope genes, p24 and cholera toxin B subunit in mice.

    abstract::No effective prophylactic or therapeutic vaccine against HIV-1 in humans is currently available. This study analyzes the immunogenicity and safety of a recombinant attenuated vaccinia virus. A chimeric gene of HIV-1 multi-epitope genes containing CpG ODN and cholera toxin B subunit (CTB) was inserted into Chinese vacc...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Du S,Wang Y,Liu C,Wang M,Zhu Y,Tan P,Ren D,Li X,Tian M,Yin R,Li C,Jin N

    更新日期:2015-07-01 00:00:00

  • Pan-serotypic detection of foot-and-mouth disease virus using a minor groove binder probe reverse transcription polymerase chain reaction assay.

    abstract::A novel assay for the pan-serotypic detection of foot-and-mouth disease virus (FMDV) was designed using a 5' conjugated minor groove binder (MGB) probe real-time RT-PCR system. This assay targets the 3D region of the FMDV genome and is capable of detecting 20 copies of a transcribed RNA standard. The linear range of t...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: McKillen J,McMenamy M,Reid SM,Duffy C,Hjertner B,King DP,Bélak S,Welsh M,Allan G

    更新日期:2011-06-01 00:00:00

  • Molecular characterization of a cDNA clone encoding the Epstein-Barr virus (EBV) DNase.

    abstract::RNA from IdUrd-treated P3HR1 cells was used for the construction of a cDNA library and screened with B95-8 EBV DNA BamHI fragment B and G probes. One clone, BG9, containing a 1.7 kb cDNA insert was further studied. Complete DNA sequence analysis revealed that BG9 encompassed the B95-8 EBV DNA sequences from nucleotide...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Chen MR,Hsu TY,Chen JY,Yang CS

    更新日期:1990-08-01 00:00:00

  • Development of a multiplex AmpliDet RNA for the simultaneous detection of Potato leafroll virus and Potato virus Y in potato tubers.

    abstract::A novel isothermal multiplex AmpliDet RNA system is described for the simultaneous amplification and detection of Potato leafroll virus (PLRV) and Potato virus Y (PVY) in seed potatoes. The risk of contamination by carry-over during diagnostic screening is eliminated by performing the reaction in a single closed tube....

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Klerks MM,Leone GO,Verbeek M,van den Heuvel JF,Schoen CD

    更新日期:2001-04-01 00:00:00

  • Characterization of a recombinant Newcastle disease virus expressing the green fluorescent protein.

    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


    authors: Engel-Herbert I,Werner O,Teifke JP,Mebatsion T,Mettenleiter TC,Römer-Oberdörfer A

    更新日期:2003-03-01 00:00:00

  • Detection and quantitation of HBV DNA in the WHO International Standard for HIV-1 RNA (NIBSC code: 97/656).

    abstract::Nucleic-acid amplification technology (NAT) assays have been implemented for HCV and HIV-1 in the United States, and many parts of Europe, Australia and Asia. Nucleic acid detection assays utilize many different technologies, and the WHO International Standards for nucleic acid tests are widely used to compare them. C...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Shyamala V,Cottrell J,Arcangel P,Madriaga D,Linnen J,Phelps B,Chien D

    更新日期:2004-06-01 00:00:00

  • Nucleotide-specific PCR for molecular virus typing.

    abstract::Nucleotide sequence studies detected a double-point mutation in the genomic RNA segment 4 (nt 871 and 872) of the persistent variant C/AA-pi of influenza C/Ann Arbor/1/50 virus. The 3'-end-points of two distinct PCR primers were positioned exactly at this genome location and thereby adjusted the priming determinant co...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Marschall M,Schuler A,Böswald C,Helten A,Hechtfischer A,Lapatschek M,Meier-Ewert H

    更新日期:1995-03-01 00:00:00

  • A one-step enzyme-linked immunosorbent assay to detect anti-CMV antibodies; development and clinical validation.

    abstract::A simple one-step ELISA to detect anti-CMV antibodies was developed. The test was based on the inhibition principle, and used anti-CMV coated microtitre plates, CMV nuclear antigen and anti-CMV Fab'-HRP conjugate; total assay time was 1.5 h. The use of Fab'-HRP conjugates improved discrimination between positive and n...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Wielaard F,Scherders J,Dagelinckx C,Gausset P

    更新日期:1987-08-01 00:00:00

  • Evaluation of a prototype HCV NS5b assay for typing strains of hepatitis C virus isolated from Tunisian haemodialysis patients.

    abstract::Hepatitis C virus (HCV) strains isolated from 68 haemodialysis Tunisian patients exhibiting chronic infection were genotyped targeting the NS5b region of the HCV genome using a prototype assay developed by Bayer HealthCare-Diagnostics (TRUGENE NS5b HCV). The overall results were compared to those obtained with another...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Othman SB,Trabelsi A,Monnet A,Bouzgarrou N,Grattard F,Beyou A,Bourlet T,Pozzetto B

    更新日期:2004-08-01 00:00:00

  • Real-time assay for quantitative detection of non-persistently transmitted Plum pox virus RNA targets in single aphids.

    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


    authors: Olmos A,Bertolini E,Gil M,Cambra M

    更新日期:2005-09-01 00:00:00

  • A novel flow cytometry-based tool for determining the efficiency of human cytomegalovirus infection in THP-1 derived macrophages.

    abstract::Human cytomegalovirus (hCMV) is a ubiquitous pathogen that causes congenital infection and severe infections in immunocompromised patients. Chronic hCMV infection may also play an important role in immunosenescence and adverse health outcomes in older adults. THP-1, a human monocytic cell line and its derived macropha...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Li H,Mao G,Carlson J,Leng SX

    更新日期:2015-09-01 00:00:00

  • Development of SYBR Green based real time PCR assay for detection of monodon baculovirus in Penaeus monodon.

    abstract::Shrimp farming is one of the most important aquaculture activities. Expansion and intensification of shrimp farming has been accompanied with the outbreak of diseases, which threaten the development and sustainability of the industry. Viral diseases are the major challenges faced by shrimp farming industries. The prev...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Ramesh Kumar D,Sanjuktha M,Rajan JJ,Ananda Bharathi R,Santiago TC,Alavandi SV,Poornima M

    更新日期:2014-09-01 00:00:00

  • Comparative analysis and generation of a robust HIV-1 DNA quantification assay.

    abstract::HIV-1 infection cannot be cured due to the presence of the latent reservoir (LR). Novel cure or treatment strategies, such as "shock and kill" or therapeutic vaccination, aim to reduce or eradicate the LR. Cure strategies utilise robust DNA quantification assays to measure the change in the LR in low copy scenarios. N...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Thomas J,Ruggiero A,Procopio FA,Pantaleo G,Paxton WA,Pollakis G

    更新日期:2019-01-01 00:00:00

  • A simple method for cloning the complete begomovirus genome using the bacteriophage phi29 DNA polymerase.

    abstract::The bacteriophage phiDNA polymerase amplifies circular DNA in a rolling circle amplification mechanism. This characteristic was applied to amplify and clone the complete circular DNA genome of a begomovirus. Total DNA extracted from infected tissue was used as the template of an amplification reaction using the commer...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Inoue-Nagata AK,Albuquerque LC,Rocha WB,Nagata T

    更新日期:2004-03-15 00:00:00

  • Determination of hepatitis C virus genotype by Pyrosequencing.

    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


    authors: Elahi E,Pourmand N,Chaung R,Rofoogaran A,Boisver J,Samimi-Rad K,Davis RW,Ronaghi M

    更新日期:2003-05-01 00:00:00

  • Evaluation of viral-lysate enzyme-linked immunosorbent assay kits for detecting human immunodeficiency virus (type 1) infections using human sera standardized by quantitative western blotting.

    abstract::The first generation of proprietary reagents for detecting antibodies to the Human Immunodeficiency Virus Type 1 (HIV-1) by enzyme-linked immunosorbent assay (ELISA) used as antigen partially purified virus from cell culture lysates. These tests, which are still in use, may vary in their antibody measurement capabilit...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Hardy CT,Damrow TA,Villareal DB,Kenny GE

    更新日期:1992-06-01 00:00:00

  • Multiplex real-time PCR for the detection and quantification of latent and persistent viral genomes in cellular or plasma blood fractions.

    abstract::In common with latent viruses such as herpesviruses, parvovirus B19, HBV and GBV-C are contained successfully by the immune response and persist in the host. When immune control breaks down, reactivation of both latent and persistent viruses occurs. Two multiplex assays were developed (B19, HBV, HHV-8), (EBV, CMV, VZV...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Compston LI,Sarkobie F,Li C,Candotti D,Opare-Sem O,Allain JP

    更新日期:2008-07-01 00:00:00