Simultaneous detection of two independent antigens by double staining with two mouse monoclonal antibodies.

Abstract:

:Simultaneous detection of two antigens by immunostaining usually requires primary antibodies from two different species or a hapten modification of one of the antibodies if they are from the same species. A novel double staining method is described for immunodetection of two independent antigens using two mouse monoclonal antibodies. The principle of the method is the following: The first antigen is detected by a monoclonal antibody that is diluted so extensively that it cannot be recognized with conventional detection systems. A highly sensitive biotin-tyramide amplification system is used to identify this antibody. The second antigen is stained with a monoclonal antibody by dilution and detected by conventional immunostaining. The method was tested for both alkaline-phosphatase staining on paraffin sections and immunofluorescence staining on cultured cells in cytospin preparation. The absence of cross-reaction in the former system was demonstrated by the mutually exclusive detection of T- and B-cells in human lymph nodes or T-cells and carcinoma cells in nasopharyngeal carcinoma biopsies. Similarly, the EBV encoded EBNA2 and ZEBRA proteins showed a mutually exclusive staining by immunofluorescence on B95-8 cells. The method could be used to demonstrate co-expression of two independent antigens in the same cells, such as PCNA and keratin in carcinoma cells in paraffin sections and for EBNA2 and LMP1 EBV proteins in immunofluorescence preparations of B95-8 cells.

journal_name

J Virol Methods

authors

Teramoto N,Szekely L,Pokrovskaja K,Hu LF,Yoshino T,Akagi T,Klein G

doi

10.1016/s0166-0934(98)00048-2

subject

Has Abstract

pub_date

1998-07-01 00:00:00

pages

89-97

issue

1

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(98)00048-2

journal_volume

73

pub_type

杂志文章
  • A flow-through cell counting assay for point-of-care enumeration of CD4 T-cells.

    abstract::CD4 T-cell count is a priority for staging HIV disease and guiding clinical management as part of HIV care. Conventional CD4 T-cell enumeration methods based on flow cytometry are expensive, require well-trained personnel, and are challenging to use in rural, resource-scarce areas. A simple CD4 T-cell count test that ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2019.05.012

    authors: Bystryak S,Bandwar RP,Santockyte R

    更新日期:2019-09-01 00:00:00

  • A multivariate statistical analysis to follow the course of disease after infection of cats with different strains of the feline immunodeficiency virus (FIV).

    abstract::A descriptive multivariate assay is described which is suitable to analyze results of a biological experiment with small sample size but high qualitative and quantitative complexity of variables. This type of assay allows evaluation of multiple variables observed in the course of an experimental virus infection (e.g. ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/s0166-0934(02)00024-1

    authors: de Monte M,Nonnenmacher H,Brignon N,Ullmann M,Martin JP

    更新日期:2002-05-16 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

    doi:10.1016/0166-0934(94)00122-w

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

    更新日期:1995-02-01 00:00:00

  • Clinical utility of 2',5'-oligoadenylate synthetase activity measurement: using whole blood as a highly sensitive method to detect the effects of IFN.

    abstract::Research progress on the pleiotropic effects of interferons (IFN) has thus far required detecting responses by weak IFN signals. The activity of 2',5'-oligoadenylate synthetase (2-5OAS) is a valuable indicator in the prognosis and IFN treatment of patients with viral diseases such as hepatitis B and C. Although serum ...

    journal_title:Journal of virological methods

    pub_type: 临床试验,杂志文章

    doi:10.1016/j.jviromet.2006.05.005

    authors: Uno K,Suginoshita Y,Kakimi K,Moriyasu Y,Nakano K,Nakamura N,Fujita T,Horino Y,Sato T,Kishida T

    更新日期:2006-09-01 00:00:00

  • The creation of stable cell lines expressing Ebola virus glycoproteins and the matrix protein VP40 and generating Ebola virus-like particles utilizing an ecdysone inducible mammalian expression system.

    abstract::Ebola virus is a filovirus that causes hemorrhagic fever in humans and is associated with case fatality rates of up to 90%. The lack of therapeutic interventions in combination with the threat of weaponizing this organism has enhanced research investigations. The expression of key viral proteins and the production of ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2007.12.004

    authors: Melito PL,Qiu X,Fernando LM,deVarennes SL,Beniac DR,Booth TF,Jones SM

    更新日期:2008-03-01 00:00:00

  • An efficient and rapid influenza gene cloning strategy for reverse genetics system.

    abstract::Influenza reverse genetics plays vital roles in understanding influenza molecular characteristics and vaccine development. However, current influenza reverse genetics heavily depends on restriction enzyme and ligation for gene cloning. The traditional cloning process of influenza eight fragments for virus rescuing gen...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2015.06.001

    authors: Shao H,Fan Z,Wan Z,Tian X,Chen H,Perez DR,Qin A,Ye J

    更新日期:2015-09-15 00:00:00

  • Detection of adenovirus by rapid 24-well plate centrifugation and conventional cell culture with dexamethasone.

    abstract::Two methods for rapid detection of adenovirus were tested: (i) 24-well plate centrifugation followed by staining with a monoclonal antibody after incubation for 24 h and 48 h, and (ii) pretreatment of A549 cells used in conventional cell culture and 24-well plate centrifugation with 10(-5)M dexamethasone. Twenty-seven...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(88)90144-9

    authors: Woods GL,Yamamoto M,Young A

    更新日期:1988-06-01 00:00:00

  • Presence of enteroviruses in recreational water in Wuhan, China.

    abstract::Contaminated recreational waters pose a public health concern, as the potential for waterborne diseases exists in water contaminated with human fecal waste. Worldwide, bacterial indicators such as Escherichia coli, enterococci, and total and fecal coliform are used as indicators of water quality. However, enteric viru...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2013.06.028

    authors: Allmann E,Pan L,Li L,Li D,Wang S,Lu Y

    更新日期:2013-11-01 00:00:00

  • Evaluation of an agglutination HIV-1 + 2 antibody assay.

    abstract::Studies have shown that an HIV (HIV-PA) agglutination assay (Serodia) for the detection of antibody to human immunodeficiency virus (HIV) can be as sensitive and as specific as enzyme-linked immunosorbent assay (ELISA). However, since this HIV assay was designed to detect antibody to the HIV-1 virus, a substantial num...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(94)00092-u

    authors: Vercauteren G,Beelaert G,van der Groen G

    更新日期:1995-01-01 00:00:00

  • Detection of human virulence signatures in H5N1.

    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

    authors: Waybright N,Petrangelo E,Lowary P,Bogan J,Mulholland N

    更新日期:2008-12-01 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

    doi:10.1016/0166-0934(83)90074-5

    authors: de Vries J,Kruining J,Heijtink RA

    更新日期:1983-02-01 00:00:00

  • Real-time PCR for the quantitation of Tomato yellow leaf curl Sardinia virus in tomato plants and in Bemisia tabaci.

    abstract::Tomato yellow leaf curl Sardinia virus (TYLCSV) (Geminiviridae) is an important pathogen severely affecting tomato production in the Mediterranean basin. Although diagnostic protocols are available for its detection in plants and its vector Bemisia tabaci (Gennadius), suitable tools for estimating and comparing viral ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2007.09.015

    authors: Mason G,Caciagli P,Accotto GP,Noris E

    更新日期:2008-02-01 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

    doi:10.1016/0166-0934(86)90056-x

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

    更新日期:1986-07-01 00:00:00

  • Viral elution and concentration method for detection of influenza A viruses in mud by real-time RT-PCR.

    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

    authors: Deboosere N,Horm SV,Delobel A,Gachet J,Buchy P,Vialette M

    更新日期:2012-01-01 00:00:00

  • Production of Ebola virus-like particles in Drosophila melanogaster Schneider 2 cells.

    abstract::In this study, we generated recombinant virus-like particles (VLPs) against family Filoviridae, genus Ebolavirus, species Zaire ebolavirus, strain Makona (EBOV) in Drosophila melanogaster Schneider 2 (S2) cells using the EBOV Makona. S2 cells were cotransfected with four viral plasmids encoding EBOV Makona proteins an...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2018.08.016

    authors: Park EM,Park SW,Lee YJ,Lee WJ,Choi W

    更新日期:2018-11-01 00:00:00

  • Cost-effective HIV-1 virological monitoring in resource-limited settings using a modified commercially available qPCR RNA assay.

    abstract::Virological monitoring through plasma viral load (PVL) quantification is essential for clinical management of HIV patients undergoing antiretroviral treatment (ART), and for detecting treatment failure. Quantitative PCR (qPCR)-based tests are the gold standard for measuring PVL. Largely because of their high cost, how...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2017.05.007

    authors: Boobalan J,Torti A,Dinesha TR,Solomon SS,Balakrishnan P,Saravanan S

    更新日期:2017-10-01 00:00:00

  • A PCR-restriction enzyme technique for determining dengue virus subgroups within serotypes.

    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

    authors: Vorndam V,Kuno G,Rosado N

    更新日期:1994-07-01 00:00:00

  • Use of hepatitis E IgG avidity for diagnosis of hepatitis E infection.

    abstract::The diagnosis of acute hepatitis E infection is based on the detection of HEV RNA or specific IgM in immunocompetent patients. Viraemia and excretion of HEV RNA in faeces are not observed in all patients and commercial kits vary in their performance for anti-HEV IgM detection. Additional diagnostic tests must therefor...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2009.11.028

    authors: Bigaillon C,Tessé S,Lagathu G,Nicand E

    更新日期:2010-03-01 00:00:00

  • Specific detection of monkeypox virus by polymerase chain reaction.

    abstract::The open reading frame coding for the A-type inclusion body protein (ATI) of monkeypox virus (MPV) was identified and sequenced for two strains. Nucleotide sequence comparison revealed 72-95.3% homology with the reported open reading frame sequences of the ATIs of other orthopoxvirus species, such as variola, vaccinia...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/s0166-0934(98)00099-8

    authors: Neubauer H,Reischl U,Ropp S,Esposito JJ,Wolf H,Meyer H

    更新日期:1998-10-01 00:00:00

  • Detection of human papillomavirus DNA and mRNA using synthetic, type-specific oligonucleotide probes.

    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

    authors: Faulkner-Jones BE,Tabrizi SN,Borg AJ,Roche PJ,Haralambidis J,Coghlan JP,Garland SM

    更新日期:1993-03-01 00:00:00

  • Rapid detection of human parvovirus B19 DNA by dot-hybridization and the polymerase chain reaction.

    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

    authors: Salimans MM,Holsappel S,van de Rijke FM,Jiwa NM,Raap AK,Weiland HT

    更新日期:1989-01-01 00:00:00

  • The selection and characterization of human monoclonal antibodies to human cytomegalovirus.

    abstract::This communication describes the application of Epstein-Barr virus lymphocyte transformation technology to the production of human monoclonal antibodies specific for human cytomegalovirus. A group of such IgG antibodies have been characterized in terms of subclass, light-chain composition, specificity for particular v...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(86)90003-0

    authors: Redmond MJ,Leyritz-Wills M,Winger L,Scraba DG

    更新日期:1986-08-01 00:00:00

  • Comparison of somatic and F+ coliphage enumeration methods with large volume surface water samples.

    abstract::Coliphages are alternative fecal indicators that may be suitable surrogates for viral pathogens, but majority of standard detection methods utilize insufficient volumes for routine detection in environmental waters. We compared three somatic and F+ coliphage methods based on a paired measurement from 1 L samples colle...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2018.08.007

    authors: McMinn BR,Rhodes ER,Huff EM,Wanjugi P,Ware MM,Nappier SP,Cyterski M,Shanks OC,Oshima K,Korajkic A

    更新日期:2018-11-01 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

    doi:10.1016/j.jviromet.2011.03.008

    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

  • 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

    doi:10.1016/j.jviromet.2012.05.003

    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

  • 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

    doi:10.1016/0166-0934(91)90108-c

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

    更新日期:1991-10-01 00:00:00

  • Real-time PCR assay to identify variants of Vaccinia virus: implications for the diagnosis of bovine vaccinia in Brazil.

    abstract::Naturally occurring infections of Vaccinia virus (VACV) have been recognized in Brazil during the past 10 years. Human Brazilian Vaccinia virus (BVV) infections typically occur as a zoonosis transferred from affected dairy cows to their handlers. Outbreaks have caused notable economic losses to the rural community in ...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2008.05.028

    authors: de Souza Trindade G,Li Y,Olson VA,Emerson G,Regnery RL,da Fonseca FG,Kroon EG,Damon I

    更新日期:2008-09-01 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

    doi:10.1016/0166-0934(85)90129-6

    authors: Katz JB,van Deusen RA

    更新日期:1985-12-01 00:00:00

  • Development of sheep kidney cells with increased resistance to different subgenotypes of BVDV-1 by RNA interference.

    abstract::Bovine viral diarrhea virus (BVDV) should be a ubiquitous viral pathogen to the cattle and sheep industry. This pathogen is responsible for severe economic losses. We previously showed that plasmid-mediated dual short hairpin RNA (shRNA) efficiently inhibit BVDV replication in bovine kidney epithelial (MDBK) cells. In...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/j.jviromet.2015.03.014

    authors: Ni W,Qiao J,Ma Q,Wang J,Wang D,Zhao X,Cao Y,Li Q,Hu S,Chen C

    更新日期:2015-06-15 00:00:00

  • Poly A-linked non-isotopic microtiter plate reverse transcriptase assay for sensitive detection of clinical human immunodeficiency virus isolates.

    abstract::A colorimetric reverse transcriptase assay (cRT assay) was developed for quantitative detection of HIV-1. In this format, reverse transcriptase incorporates biotin-labeled dUTP onto oligo-dT primers hybridized to poly A templates. The templates are covalently bound to the surface of microtiter wells. The amount of inc...

    journal_title:Journal of virological methods

    pub_type: 杂志文章

    doi:10.1016/0166-0934(95)00073-5

    authors: Suzuki K,Saito T,Kondo M,Osanai M,Watanabe S,Kano T,Kano K,Imai M

    更新日期:1995-11-01 00:00:00