Development of a real-time RT-PCR assay for detection of resistance to oseltamivir in influenza A pandemic (H1N1) 2009 virus using single nucleotide polymorphism probes.


:Resistance to oseltamivir in pandemic (H1N1) 2009 influenza A virus is linked to an amino acid change from histidine (H) to tyrosine (Y) at position 275 in the neuraminidase protein (NA). A real-time one step RT-PCR assay using single nucleotide polymorphism (SNP) probes was developed to detect this mutation in respiratory specimens. The limit of detection was 47.6 copies/reaction for wild-type H275 RNA and 52.9 copies/reaction for the mutant H275Y RNA. The assay did not cross-react with other respiratory pathogens. The clinical sensitivity and specificity of the assay was compared to the gold standard Sanger sequencing method using 25 sensitive, 15 resistant and 20 negative samples. The sensitivity and specificity was 88.0% and 100% respectively with the SOIV_Osel_SEN probe designed to detect the H275 allele and 100% for the SOIV_Osel_RES probe detecting the 275Y allele. The sensitivity of the assay using nine admixtures of sensitive and resistant alleles was 88.9% and 77.8% with the SOIV_Osel_SEN probe and SOIV_Osel_RES probe respectively. The presence of mixed sensitive and resistant alleles in patient samples and mixtures of in vitro RNA were detected reproducibly. This assay can be used for screening of original samples for oseltamivir resistance without the need for culture and phenotypic testing.


J Virol Methods


Wong S,Pabbaraju K,Wong A,Fonseca K,Drews SJ




Has Abstract


2011-05-01 00:00:00














  • Detection of Epstein--Barr virus-determined nuclear antigen-2 mRNA by in situ hybridization.

    abstract::A mRNA in situ hybridization method was developed to detect mRNA of Epstein--Barr virus-determined nuclear antigen-2 (EBNA2). Strong in situ hybridization signals were detected in EBNA2-transfected cells with the antisense probe but not with the sense probe of this mRNA. Hybridization signals were also found in formal...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Shimakage M,Sasagawa T

    更新日期:2001-04-01 00:00:00

  • Construction, characterization and immunogenicity of a glycoprotein E negative bovine herpesvirus-1.1 Egyptian strain "Abu-Hammad".

    abstract::A full glycoprotein E (gE) deletion was generated in genome of the Egyptian BoHV-1.1 Abu-Hammad strain. Integrity of the gE negative (gE(-)) mutant virus was proved by successful specific PCR amplifications of gB, gC, tk, gD, gI and gE genes along with definite immune reaction to polyclonal anti-BoHV-1 antibody in inf...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: El-Kholy AA,Rady DI,Abdou ER,Elseafy MM,Abdelrahman KA,Soliman H

    更新日期:2013-12-01 00:00:00

  • Infectivity assays for the Autographa californica nuclear polyhedrosis virus using Spodoptera littoralis cells.

    abstract::Tissue culture ID50 and plaque assays for the Autographa californica nuclear polyhedrosis virus, using the Spodoptera littoralis cell line HPB-SL, were developed. Direct comparison using these assay methods showed that these cells were as susceptible to infection as the more commonly used Spodoptera frugiperda cell li...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Roberts PL

    更新日期:1985-01-01 00:00:00

  • Selection and characterization of single-chain recombinant antibodies against infectious haematopoietic necrosis virus from mouse phage display library.

    abstract::Six single-chain fragment variable (scFv) antibodies against infectious haematopoietic necrosis virus (IHNV) were selected from an antibody phage display library by phage display technology. The soluble scFv antibodies showed a molecular weight 32kDa by Western blot. Dot blot analysis revealed that the six scFv antibo...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Liu H,Zheng X,Shi X,Yu L,Jia P,Wang J,He J,Lan W,Liu H,Wu Z

    更新日期:2014-09-01 00:00:00

  • Oligonucleotide microarray design for detection and serotyping of human respiratory adenoviruses by using a virtual amplicon retrieval software.

    abstract::The rapid development of new molecular biology methods has improved infectious disease diagnosis, which is increasingly important to clinical management and public health. A wide variety of new methods which are more specific, sensitive and robust, such as combination of PCR and microarray technology, has gradually re...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: López-Campos G,Coiras M,Sánchez-Merino JP,López-Huertas MR,Spiteri I,Martín-Sánchez F,Pérez-Breña P

    更新日期:2007-11-01 00:00:00

  • Mapping of epitopes of VP2 protein of chicken anemia virus using monoclonal antibodies.

    abstract::To map the epitopes of VP2 protein of chicken anemia virus (CAV), VP2 was expressed as a fusion protein in Escherichia coli BL21 (DE3). The Western blot demonstrated that recombinant VP2 protein could be recognized by sera of chickens infected with CAV. Female BALB/c mice were immunized with purified recombinant VP2 p...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Wang X,Gao H,Gao Y,Fu C,Wang Z,Lu G,Cheng Y,Wang X

    更新日期:2007-08-01 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

  • Molecular detection of varicella zoster virus while keeping an eye on the budget.

    abstract::Varicella zoster virus (VZV) PCR is highly sensitive compared to traditional detection methods like culture and direct fluorescent antibody testing (DFA); however, the high cost of commercial assays prohibits their use in many clinical laboratories. Major contributors to cost are the nucleic acid extraction and the PC...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Binkhamis K,Al-Siyabi T,Heinstein C,Hatchette TF,LeBlanc JJ

    更新日期:2014-06-01 00:00:00

  • A novel method for characterizing the multi-functional C-terminal domain of the Hepadnavirus core protein.

    abstract::The Hepadnavirus core protein is a viral structural protein with an N-terminal self-assembling domain and a C-terminal protamine-like arginine-rich domain (ARD). The ARD contains four clusters of arginine residues involved in RNA binding, genome DNA synthesis, and nuclear localization. Characterization of the multi-fu...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Lu L,Liu W,Yang X

    更新日期:2009-06-01 00:00:00

  • Amplification of complete gag gene sequences from geographically distinct equine infectious anemia virus isolates.

    abstract::In the current study, primers described previously and modified versions of these primers were evaluated for amplification of full-length gag genes from different equine infectious anemia virus (EIAV) strains from several countries, including the USA, Germany and Japan. Each strain was inoculated into a primary horse ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Boldbaatar B,Bazartseren T,Koba R,Murakami H,Oguma K,Murakami K,Sentsui H

    更新日期:2013-04-01 00:00:00

  • Evaluation of a semi-automatic radioimmunoassay for hepatitis B surface antigen (HBsAg).

    abstract::The recently developed semi-automatic Hepatube system was evaluated in comparison to another radioimmunoassay for the detection of hepatitis B surface antigen (HBsAg), the manual Ausria II-125 test. After incubation of serum in anti-HBs coated tubes, the Hepatube system uses a machine to wash the tubes and to add trac...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: de Vries J,Kruining J,Heijtink RA

    更新日期:1983-02-01 00:00:00

  • Development of reverse transcription loop-mediated isothermal amplification assay for rapid detection of an emerging potyvirus: tomato necrotic stunt virus.

    abstract::Tomato necrotic stunt virus (ToNStV) is an emerging potyvirus that causes severe stunting to infected tomato plants. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for sensitive detection of ToNStV. The sensitivity of RT-LAMP was comparable to that of conventional RT-PCR, ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Li R,Ling KS

    更新日期:2014-05-01 00:00:00

  • A preliminary evaluation of five HIV antigen detection assays.

    abstract::Five commercial assays for detecting HIV antigen were evaluated using a panel of 40 coded samples in six laboratories. All assays were capable of detecting HIV-1 antigen (HIV-1 Ag), and are likely to prove useful for monitoring supernatant fluids from a variety of cell cultures. The concentration of HIV-1 protein whic...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Healey DS,Maskill WJ,Neate EV,Beaton F,Gust ID

    更新日期:1988-06-01 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

  • Antigenic analysis of intraepidemic variants of influenza A (H3N2) viruses by hyperimmune rat antisera.

    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


    authors: Pyhälä R,Pyhälä L

    更新日期:1987-03-01 00:00:00

  • The persistent infection of a canine thymus cell line by equine infectious anaemia virus and preliminary data on the production of viral antigens.

    abstract::Equine infectious anaemia virus (EIAV) was adapted to the Cf2Th cell line, a heterologous malignant line from canine thymus. A persistent infection was monitored for 100 serial passages by demonstrating the presence of virus and viral antigens at each 10th passage by electron-microscopy, immunodiffusion and immunofluo...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Bouillant AM,Nielsen K,Ruckerbauer GM,Samagh BS,Hare WC

    更新日期:1986-07-01 00:00:00

  • Identification of simian varicella virus gene 21 promoter region using green fluorescent protein.

    abstract::Clinical, pathological, immunological and virological features of simian varicella virus (SVV) infection in primates closely resemble those of varicella zoster virus (VZV) infection in humans. In ganglia infected latently of humans and monkeys, gene 21 of VZV and SVV is transcribed, respectively. We determined the nuc...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Mahalingam R,Wellish M,White T,Gilden DH

    更新日期:2000-04-01 00:00:00

  • Cytotoxicity associated with prolonged room temperature storage of serum and proposed methods for reduction of cytotoxicity.

    abstract::Canine serum preserved at room temperature (25°C) for longer than 24h is known to exhibit significant cytotoxicity. This phenomenon is one of the major reasons for the failure of virus neutralization tests. In this study, a method for reducing this cytotoxicity was investigated by applying several treatments to dog, c...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Shiraishi R,Hirayama N

    更新日期:2015-12-01 00:00:00

  • Rapid bioassay of human interferon by direct enzyme immunoassay of encephalomyocarditis virus in HEp-2 cell monolayers after a single cycle of infection.

    abstract::Multiplication of encephalomyocarditis virus (EMCV) in human HEp-2 cells, and its suppression by interferon (IFN), was demonstrated by direct enzyme immunoassay (EIA) in cell culture. EMCV was detected in glutaraldehyde fixed HEp-2 cell monolayers, in wells of 96-well plates, with a horse radish peroxidase (HRPO) labe...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Vlaspolder F,Donkers E,Harmsen T,Kraaijeveld CA,Snippe H

    更新日期:1989-04-01 00:00:00

  • Virus isolation and full-length genome sequencing of a representative canine distemper virus wild type strain of the South America 2 clade.

    abstract::Canine Distemper Virus (CDV) is a highly contagious pathogen of dogs that causes severe respiratory, gastrointestinal and nervous signs. Although vaccines have been used to prevent infections, CDV has been reported worldwide, even in vaccinated animals. In the present study, a representative wild type CDV strain (Arg2...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Romanutti C,Keller L,La Torre J,Panzera Y,Fuques E,Pérez R,Gallo Calderon M

    更新日期:2020-05-01 00:00:00

  • Enhancing the sensitivity of Dengue virus serotype detection by RT-PCR among infected children in India.

    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


    authors: Ahamed SF,Vivek R,Kotabagi S,Nayak K,Chandele A,Kaja MK,Shet A

    更新日期:2017-06-01 00:00:00

  • Performance evaluation of direct fluorescent antibody, Focus Diagnostics Simplexa™ Flu A/B & RSV and multi-parameter customized respiratory Taqman® array card in immunocompromised patients.

    abstract:BACKGROUND:Molecular assays for diagnosis of Flu A, Flu B, and RSV with short turn-around-time (TAT) are of considerable clinical importance. In addition, rapid and accurate diagnosis of a large panel of viral and atypical pathogens can be crucial in immunocompromised patients. OBJECTIVES:First, to evaluate the perfor...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Steensels D,Reynders M,Descheemaeker P,Curran MD,Jacobs F,Denis O,Delforge ML,Montesinos I

    更新日期:2017-07-01 00:00:00

  • Rapid confirmation by RFLP of transfer to vaccine candidate reassortment viruses of the principal 'high yield' gene of influenza A viruses.

    abstract::Influenza vaccines must be revised constantly on almost a yearly basis because of the sequential mutations (antigenic drift) that occur as the virus responds to immunologic pressure. New, high yield (hy) reassortant viruses have proved essential to meet production needs for the supply of new vaccines. We have devised ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Brett I,Werber J,Kilbourne ED

    更新日期:2002-02-01 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

  • Comparison of conventional RT-PCR, reverse-transcription loop-mediated isothermal amplification, and SYBR green I-based real-time RT-PCR in the rapid detection of bovine viral diarrhea virus nucleotide in contaminated commercial bovine sera batches.

    abstract::Bovine viral diarrhea virus (BVDV) can contaminate biological products produced in bovine or porcine cells or manufactured using bovine sera. A rapid, specific, sensitive, and practical method of detecting BVDV in bio-products is needed. The purpose of this study was to compare three assays with respect to their abili...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Zhang SQ,Tan B,Li P,Wang FX,Guo L,Yang Y,Sun N,Zhu HW,Wen YJ,Cheng SP

    更新日期:2014-10-01 00:00:00

  • Generation of dried tube specimen for HIV-1 viral load proficiency test panels: a cost-effective alternative for external quality assessment programs.

    abstract::Participation in external quality assessment programs is critical to ensure quality clinical laboratory testing. Commercially available proficiency test panels for HIV-1 virus load testing that are used commonly in external quality assessment programs remain a financial obstacle to resource-limited countries. Maintain...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Ramos A,Nguyen S,Garcia A,Subbarao S,Nkengasong JN,Ellenberger D

    更新日期:2013-03-01 00:00:00

  • A RT-PCR assay for the differential diagnosis of vesicular viral diseases of swine.

    abstract::A RT-PCR assay based on specific amplification of RNA sequences from each of the etiological agents of three important vesicular diseases that affect swine, foot-and-mouth disease virus (FMDV), swine vesicular disease virus (SVDV), and vesicular stomatitis virus (VSV), was developed. Genotype-specific primers that amp...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Núñez JI,Blanco E,Hernández T,Gómez-Tejedor C,Martín MJ,Dopazo J,Sobrino F

    更新日期:1998-06-01 00:00:00

  • DNA: DNA hybridization method for the diagnosis of hepatitis B infection.

    abstract::Hepatitis B viral (HBV) DNA was detected in a hepatoma cell line which produces hepatitis B surface antigen (HBsAg) and in patients with acute hepatitis B. The serum of one patient with acute hepatitis B was found to be infectious when injected i.v. into a chimpanzee up to a dilution of 10(-8). Hepatitis B surface ant...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Feinman SV,Berris B,Guha A,Sooknanan R,Bradley DW,Bond WW,Maynard JE

    更新日期:1984-05-01 00:00:00

  • Increased detection of human immunodeficiency virus antigen carriers by simultaneous assay of plasma and extracts from resting and phytohaemagglutinin-stimulated peripheral blood mononuclear cells.

    abstract::Antigenaemia due to human immunodeficiency virus (HIV) is thought to be significant either before the appearance of a specific antibody response, or after its decline during terminal stages. In order to increase the rate of detection of HIV antigen carriers, regardless of the stage or despite the presence of specific ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章


    authors: Ruiz-Arguelles A,Ponce-de-Leon S,Presno-Bernal M,Ponce-de-Leon A,Ortiz-Lopez R,Rojas G

    更新日期:1988-12-01 00:00:00

  • Using RT-PCR and bDNA assays to measure non-clade B HIV-1 subtype RNA.

    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


    authors: Pasquier C,Sandres K,Salama G,Puel J,Izopet J

    更新日期:1999-08-01 00:00:00