scholarly journals Detection of Norwalk Virus and Other Genogroup 1 Human Caliciviruses by a Monoclonal Antibody, RecombinantAntigen-Based Immunoglobulin M Capture Enzyme Immunoassay

1998 ◽  
Vol 36 (4) ◽  
pp. 1064-1069 ◽  
Author(s):  
James P. Brinker ◽  
Neil R. Blacklow ◽  
Mary K. Estes ◽  
Christine L. Moe ◽  
Kellogg J. Schwab ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Acute Phase ◽  
Recombinant Antigen ◽  
Immunoglobulin M ◽  
Norwalk Virus ◽  
Healthy Individuals ◽  
Test Specificity ◽  
Stool Samples ◽  
Adult Volunteers

Sera obtained from two groups of adult volunteers infected with Norwalk virus (NV) and two groups of patients involved in two natural outbreaks were tested for NV-reactive immunoglobulin M (IgM) by use of a monoclonal antibody, recombinant-antigen-based IgM capture enzyme immunoassay (EIA). No NV-reactive IgM was detected in the preinoculation sera of 15 volunteers, and 14 of 15 showed NV-reactive antibodies postinfection with NV. All of the volunteers showed IgG seroconversion to NV. In the outbreak studies, all 9 persons in one outbreak and 19 of 24 in another outbreak had NV-reactive IgM. In the first outbreak, only three of nine seroconverted to NV, which was likely due to late collection of acute-phase sera. In the second outbreak, 21 of 24 showed IgG seroconversion to NV. Sequencing of viruses isolated from five stool samples selected from those in the second outbreak showed that they were human calicivirus (HuCV) genogroup 1 viruses related, but not identical, to NV. In the volunteer studies, NV-reactive IgM was first detected 8 days postinoculation. The time of development of NV-reactive IgM antibodies in natural outbreaks was estimated to be similar to that found in the volunteer studies. Sera from three Hawaii virus-infected volunteers, four Snow Mountain virus patients, and 80 healthy individuals were negative for NV-reactive IgM, indicating test specificity for HuCV genogroup I infections. This capture IgM EIA is suitable for diagnosis of NV and other HuCV genogroup I infections and is especially useful when sera and fecal samples have not been collected early in the course of an outbreak.

scholarly journals Immunoglobulin M Antibody Test To Detect Genogroup II Norwalk-Like Virus Infection

1999 ◽  
Vol 37 (9) ◽  
pp. 2983-2986 ◽  
Author(s):  
James P. Brinker ◽  
Neil R. Blacklow ◽  
Xi Jiang ◽  
Mary K. Estes ◽  
Christine L. Moe ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Virus Infection ◽  
Antibody Test ◽  
Virus Antigen ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Norwalk Virus ◽  
Viral Etiology ◽  
Specific Immunoglobulin ◽  
Adult Volunteers

Sera obtained from adult volunteers inoculated with genogroup II Norwalk-like viruses (NLVs), Hawaii virus, and Snow Mountain virus and from patients involved in outbreaks of gastroenteritis were tested for genogroup II NLV Mexico virus-specific immunoglobulin M (IgM) by use of a monoclonal antibody, recombinant Mexico virus antigen (rMXV)-based IgM capture enzyme-linked immunosorbent assay (ELISA). Sera from genogroup I Norwalk virus (NV)-inoculated volunteers and from patients involved in a genogroup I NLV outbreak were also tested. In sera from those infected with genogroup I NV or NLVs in volunteer and outbreak studies, only 3 of 25 were rMXV IgM positive; in contrast, 24 of 25 were IgM positive for recombinant NV (rNV). In sera from those infected with genogroup II NLVs in volunteer and outbreak studies, 28 of 47 were rMXV IgM positive and none were IgM positive for rNV, showing the specificity of each IgM test for its respective genogroup. In an outbreak of gastroenteritis not characterized as being of viral etiology but suspected to be due to NV, 7 of 13 persons had IgM responses to rMXV, whereas none had IgM responses to rNV, thus establishing the diagnosis as genogroup II NLV infection. The rMXV-based IgM capture ELISA developed is specific for the diagnosis of genogroup II NLV infections.

scholarly journals Diagnosis of Norwalk Virus Infection by Indirect Enzyme Immunoassay Detection of Salivary Antibodies to Recombinant Norwalk Virus Antigen

2004 ◽  
Vol 11 (6) ◽  
pp. 1028-1034 ◽  
Author(s):  
Christine L. Moe ◽  
Arnie Sair ◽  
Lisa Lindesmith ◽  
Mary K. Estes ◽  
Lee-Ann Jaykus
Keyword(s):  
Enzyme Immunoassay ◽  
Fold Increase ◽  
Virus Antigen ◽  
Attractive Alternative ◽  
Norwalk Virus ◽  
Serum Samples ◽  
Salivary Iga ◽  
Mucosal Antibody ◽  
Antibody Assays ◽  
Stool Samples

ABSTRACT Simple diagnostic tests are needed for the detection of norovirus (NoV) outbreaks. Salivary antibody assays provide an attractive alternative to collecting and testing serum or stool samples. Antibodies to Norwalk virus (NV) in oral fluid samples were compared with NV antibodies in serum collected from 38 volunteers challenged with NV inoculum. Pre- and postchallenge (day 4, 8, 14, and 21) saliva and serum samples were examined by enzyme immunoassay (EIA) using recombinant NV antigen. Of 18 infected subjects (those who shed NV in stool or who demonstrated immunoglobulin G [IgG] seroconversion), 15 (83%) had ≥4-fold increases in NV-specific salivary IgA and 15 (83%) had ≥4-fold increases in NV-specific salivary IgG when prechallenge and postchallenge saliva samples were compared. When the results of the IgA and IgG assays were combined, all 18 infected subjects showed ≥4-fold increases in NV-specific salivary IgG or IgA postchallenge titers compared to their prechallenge titers. One of 19 uninfected subjects had a ≥4-fold increase in NV-specific salivary IgG. The sensitivity of the combined assay results was 100%, and the specificity was 95%. NV-specific salivary IgA titers peaked around 14 days postchallenge. NV-specific salivary IgG and serum IgG titers continued to rise through 21 days postchallenge. The application of this EIA to an elementary school outbreak indicated that 67% of the subjects with confirmed infections had >4-fold rises in anti-NoV IgA when an antigen in the same genetic cluster as the outbreak virus was used. This is the first documented mucosal antibody response to NoV in children. This EIA provides a useful approach for diagnosing NoV outbreaks.

scholarly journals Analysis of a Routinely Used Commercial Anti-Chikungunya IgM ELISA Reveals Cross-Reactivities with Dengue in Brazil: A New Challenge for Differential Diagnosis?

Diagnostics ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 819
Author(s):  
Monique da Rocha Queiroz Lima ◽  
Raquel Curtinhas de Lima ◽  
Elzinandes Leal de Azeredo ◽  
Flavia Barreto dos Santos
Keyword(s):  
Disease Surveillance ◽  
Endemic Area ◽  
Chikungunya Virus ◽  
Elisa Test ◽  
Cross Reactivity ◽  
Immunoglobulin M ◽  
Diagnostic Tools ◽  
Healthy Individuals ◽  
Denv Serotypes ◽  
Igm Elisa

In Brazil, chikungunya emerged in 2014, and by 2016, co-circulated with other arbovirosis, such as dengue and zika. ELISAs (Enzyme-Linked Immunosorbent Assays) are the most widely used approach for arboviruses diagnosis. However, some limitations include antibody cross reactivities when viruses belong to the same genus, and sensitivity variations in distinct epidemiological scenarios. As chikungunya virus (CHIKV) is an alphavirus, no serological cross reactivity with dengue virus (DENV) should be observed. Here, we evaluated a routinely used chikungunya commercial IgM (Immunoglobulin M) ELISA test (Anti-Chikungunya IgM ELISA, Euroimmun) to assess its performance in confirming chikungunya in a dengue endemic area. Samples (n = 340) representative of all four DENV serotypes, healthy individuals and controls were tested. The Anti-CHIKV IgM ELISA test had a sensitivity of 100% and a specificity of 25.3% due to the cross reactivities observed with dengue. In dengue acute cases, the chikungunya test showed an overall cross-reactivity of 31.6%, with a higher cross-reactivity with DENV-4. In dengue IgM positive cases, the assay showed a cross-reactivity of 46.7%. Serological diagnosis may be challenging and, despite the results observed here, more evaluations shall be performed. Because distinct arboviruses co-circulate in Brazil, reliable diagnostic tools are essential for disease surveillance and patient management.

scholarly journals A Novel Mouse Monoclonal Antibody C42 against C-Terminal Peptide of Alpha-1-Antitrypsin

2021 ◽  
Vol 22 (4) ◽  
pp. 2141
Author(s):  
Srinu Tumpara ◽  
Elena Korenbaum ◽  
Mark Kühnel ◽  
Danny Jonigk ◽  
Beata Olejnicka ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometry ◽  
Western Blotting ◽  
Complex Mixture ◽  
Immunoglobulin M ◽  
Confocal Laser ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot

The C-terminal-fragments of alpha1-antitrypsin (AAT) have been identified and their diverse biological roles have been reported in vitro and in vivo. These findings prompted us to develop a monoclonal antibody that specifically recognizes C-36 peptide (corresponding to residues 359–394) resulting from the protease-associated cleavage of AAT. The C-36-targeting mouse monoclonal Immunoglobulin M (IgM) antibody (containing κ light chains, clone C42) was generated and enzyme-linked immunosorbent assay (ELISA)-tested by Davids Biotechnologie GmbH, Germany. Here, we addressed the effectiveness of the novel C42 antibody in different immunoassay formats, such as dot- and Western blotting, confocal laser microscopy, and flow cytometry. According to the dot-blot results, our novel C42 antibody detects the C-36 peptide at a range of 0.1–0.05 µg and shows no cross-reactivity with native, polymerized, or oxidized forms of full-length AAT, the AAT-elastase complex mixture, as well as with shorter C-terminal fragments of AAT. However, the C42 antibody does not detect denatured peptide in SDS-PAGE/Western blotting assays. On the other hand, our C42 antibody, unconjugated as well as conjugated to DyLight488 fluorophore, when applied for immunofluorescence microscopy and flow cytometry assays, specifically detected the C-36 peptide in human blood cells. Altogether, we demonstrate that our novel C42 antibody successfully recognizes the C-36 peptide of AAT in a number of immunoassays and has potential to become an important tool in AAT-related studies.

Development of a Monoclonal Antibody-Based Enzyme Immunoassay for the Pyrethroid Insecticide Deltamethrin

2010 ◽  
Vol 58 (14) ◽  
pp. 8189-8195 ◽  
Author(s):  
Ye Kong ◽  
Qi Zhang ◽  
Wen Zhang ◽  
Shirley J. Gee ◽  
Peiwu Li
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Pyrethroid Insecticide

scholarly journals Measurement of IgA responses following Norwalk virus infection and other human caliciviruses using a recombinant Norwalk virus protein EIA

1994 ◽  
Vol 113 (1) ◽  
pp. 143-151 ◽  
Author(s):  
S. P. Parker ◽  
W. D. Cubitt
Keyword(s):  
Virus Infection ◽  
Enzyme Immunoassay ◽  
Capsid Protein ◽  
Causative Agent ◽  
Food Poisoning ◽  
Hospital Staff ◽  
Virus Protein ◽  
Norwalk Virus ◽  
Major Outbreak ◽  
Virus Capsid Protein

SUMMARYAn enzyme immunoassay employing recombinant Norwalk virus capsid protein was evaluated for the measurement of IgA responses. Tests on 23 volunteers and patients known to have been infected with Norwalk virus (NV) showed that 19 developed significant IgA responses, 2 had unchanging levels of IgA and 2 failed to respond. There was no evidence of IgA responses to NV following infection with Hawaii or Snow Mountain-like viruses.Tests on sera from patients involved in outbreaks associated with eating contaminated shellfish suggest that some patients may have been infected with more than one strain of calicivirus. The use of the rNV EIA for measuring IgA and IgG responses in patients involved in a major outbreak of food poisoning affecting hospital staff indicated that the causative agent was probably NV.

scholarly journals Bound/free separation methods in steroid enzyme immunoassay with monoclonal antibody.

1988 ◽  
Vol 36 (9) ◽  
pp. 3525-3531 ◽  
Author(s):  
HIROSHI HOSODA ◽  
REIKO TSUKAMOTO ◽  
SAKIKO TAMURA ◽  
TOSHIO NAMBARA
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay
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