scholarly journals Evaluation of a Commercially Available Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay Kit for Diagnosing Acute Dengue Infections

1999 ◽  
Vol 6 (5) ◽  
pp. 741-744 ◽  
Author(s):  
Kevin R. Porter ◽  
Susana Widjaja ◽  
Handinata Darmawan ◽  
Lohita ◽  
Sri Hartati Hadiwijaya ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Igm Antibodies ◽  
Paired Samples ◽  
Secondary Infections ◽  
Test Kit ◽  
Antibody Capture ◽  
Negative Controls

ABSTRACT Recently, commercially available kits for the detection of anti-dengue virus (anti-DEN) immunoglobulin M (IgM) antibodies have been developed. These standardized assays have greatly enhanced our ability to effectively diagnose DEN infections. We conducted an evaluation of a test kit manufactured by MRL Diagnostics Inc. that is designed to detect anti-DEN IgM antibodies. Eighty paired samples from DEN-infected individuals were tested by the MRL DEN Fever Virus IgM Capture enzyme-linked immunosorbent assay (ELISA), the PanBio Duo ELISA, the PanBio Rapid Immunochromatographic Test (PRIT), and the IgM-IgG antibody capture (MAC/GAC) ELISA. All infections were confirmed by either PCR-assisted detection of DEN transcripts or by DEN isolation in C6/36 cells. Seventeen paired samples from individuals with no evidence of acute DEN infection were used as negative controls. The PRIT had the best sensitivity (100%), whereas the MAC/GAC ELISA and the PanBio Duo assay had the highest levels of specificity. The MRL ELISA and the PanBio Duo assay were the top performers when taking into consideration both sensitivity and specificity. All assays were able to detect DEN-specific antibodies in samples from patients with either primary or secondary infections, regardless of the infecting DEN serotype.

scholarly journals Evaluation of an Immunoglobulin M Capture Enzyme-Linked Immunosorbent Assay for Diagnosis of Orientia tsutsugamushi Infection

2003 ◽  
Vol 10 (3) ◽  
pp. 394-398 ◽  
Author(s):  
Won-Jong Jang ◽  
Myung-Suk Huh ◽  
Kyung-Hee Park ◽  
Myung-Sik Choi ◽  
Ik-Sang Kim
Keyword(s):  
Immunosorbent Assay ◽  
Scrub Typhus ◽  
Microtiter Plate ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Orientia Tsutsugamushi ◽  
Serum Samples ◽  
Capture Elisa ◽  
Igm Antibodies ◽  
Immunodominant Antigen

ABSTRACT To differentiate scrub typhus from other acute febrile diseases, a rapid and reliable serological diagnosis is important. We developed an immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (ELISA) for diagnosis of recent Orientia tsutsugamushi infections in humans. The 56-kDa major outer membrane protein of O. tsutsugamushi is well known as the most immunodominant antigen in scrub typhus. The test is based on the use of the biotinylated recombinant 56-kDa protein of O. tsutsugamushi Boryong, Bor56, which was expressed as a fusion protein with a maltose-binding protein in Escherichia coli. In the test, the serum IgM antibodies were captured by anti-human IgM antibodies coated onto a microtiter plate. The captured IgM antibodies were revealed through sequential addition of biotinylated Bor56 antigen and peroxidase-conjugated streptavidin to the plate. The IgM capture ELISA was compared with the immunofluorescence antibody assay (IFA) by testing 176 serum samples from patients with diagnosed cases of rickettsial disease and patients with other acute febrile diseases. Of the 81 IgG IFA-positive samples, 78 tested positive (sensitivity, 96.3%) and all 31 IgM IFA-positive samples tested positive (sensitivity, 100%) by the IgM capture ELISA. The specificity of the IgM capture ELISA was 99%, and 1 of the 95 IFA-negative samples was positive in the assay. These results strongly suggest that IgM capture ELISA using the recombinant Bor56 antigen is a reliable and detailed method for the detection of early O. tsutsugamushi infection.

scholarly journals Peptide-Based OspC Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Lyme Borreliosis

1998 ◽  
Vol 36 (12) ◽  
pp. 3474-3479 ◽  
Author(s):  
Marianne J. Mathiesen ◽  
Michael Christiansen ◽  
Klaus Hansen ◽  
Arne Holm ◽  
Eva Åsbrink ◽  
...  
Keyword(s):  
Lyme Borreliosis ◽  
Immunosorbent Assay ◽  
Erythema Migrans ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Amino Acid Residues ◽  
Serum Samples ◽  
Igg Antibody ◽  
Acrodermatitis Chronica Atrophicans ◽  
Increased Sensitivity

Sera from 210 patients with Lyme borreliosis (LB) were studied by an enzyme-linked immunosorbent assay (ELISA) based on a synthetic peptide (pepC10) comprising the C-terminal 10-amino-acid residues of OspC of Borrelia burgdorferi. We found that 36.3 and 45.0% of the serum samples from patients with erythema migrans (EM) and neuroborreliosis (NB), respectively, displayed immunoglobulin M (IgM) anti-pepC10 reactivities, while these samples rarely (≤8%) displayed IgG antibody reactivities. Sera from patients with acrodermatitis chronica atrophicans did not contain anti-pepC10 antibodies. The diagnostic performance of this newly developed peptide ELISA was compared with those of an ELISA based on the full-length recombinant OspC protein (rOspC) and a commercially available ELISA based on theB. burgdorferi flagellum (Fla). The sensitivity of the IgM pepC10 ELISA was slightly lower (P < 0.04) than that of the rOspC ELISA for EM patients (36.3 versus 43.8%), while there was no difference for NB patients (45.0 versus 48.0%). However, the optical density values obtained by the pepC10 ELISA were generally higher than those obtained by the rOspC ELISA, leading to a significantly better quantitative discrimination between seropositive patients with NB and controls (P < 0.008). The specificity of the pepC10 ELISA was similar to those of the rOspC ELISA and the Fla ELISA for relevant controls including patients with syphilis and mononucleosis. Although the overall diagnostic sensitivity of the Fla ELISA was superior, 8.8 and 12.0% of the EM and NB patients, respectively, were antibody positive only by the pepC10 ELISA. Thus, use of a diagnostic test for LB based on the detection of IgM antibodies to pepC10 and Fla has increased sensitivity for the diagnosis of early LB.

scholarly journals Development of an Immunoglobulin M (IgM) Capture Enzyme-Linked Immunosorbent Assay for Detection of Equine and Swine IgM Antibodies to Vesicular Stomatitis Virus

2001 ◽  
Vol 8 (3) ◽  
pp. 475-481 ◽  
Author(s):  
En-min Zhou ◽  
Jose Riva ◽  
Alfonso Clavijo
Keyword(s):  
Vesicular Stomatitis Virus ◽  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Competitive Elisa ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igm Antibodies ◽  
Vesicular Stomatitis ◽  
Elisa Plate

ABSTRACT An immunoglobulin M (IgM) capture enzyme-linked immunosorbent assay (MC-ELISA) was developed for the detection of primary infection of vesicular stomatitis virus (VSV) in equine and swine sera. The test was based on the use of biotinylated sheep antibodies against equine or swine IgM molecules bound to a streptavidin-coated ELISA plate. The captured IgM antibodies were detected by application of antigens prepared from the New Jersey and the Indiana VSV serotypes (VSV-NJ and VSV-IN, respectively) and mouse polyclonal antibodies against VSV-NJ and VSV-IN. The MC-ELISA was compared to a competitive ELISA (C-ELISA) and the standard microtiter serum neutralization (MTSN) assay by testing serum samples from horses and pigs experimentally infected with VSV-NJ or VSV-IN. The MC-ELISA detected specific homologous IgM antibodies from equine and swine sera as early as 5 and 4 days postinfection (DPI), respectively, and as late as 35 DPI. The MTSN test also detected antibodies as early as 5 DPI and as late as 160 DPI. In a similar fashion, the C-ELISA detected antibodies from 6 to 7 DPI and as late as 160 DPI. These results demonstrated that the MC-ELISA is a useful test for serodiagnosis of primary VSV infection in horses and pigs.

scholarly journals Comparison of Capture Immunoglobulin M (IgM) and IgG Enzyme-Linked Immunosorbent Assay (ELISA) and Nonstructural Protein NS1 Serotype-Specific IgG ELISA for Differentiation of Primary and Secondary Dengue Virus Infections

2003 ◽  
Vol 10 (4) ◽  
pp. 622-630 ◽  
Author(s):  
Pei-Yun Shu ◽  
Li-Kuang Chen ◽  
Shu-Fen Chang ◽  
Yi-Yun Yueh ◽  
Ling Chow ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Immunosorbent Assay ◽  
Primary Infection ◽  
Nonstructural Protein ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Igg Antibody ◽  
Previous Infection ◽  
Specific Igg

ABSTRACT We have found that NS1 serotype-specific immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) can be used to differentiate primary and secondary dengue virus infections. This is due to the fact that the NS1-specific IgG antibody cannot be detected before day 9 of illness for primary infection, so the NS1-specific IgG antibodies measured in acute-phase sera must come from previous infection. Comparison of NS1 serotype-specific IgG ELISA with envelope- and membrane-specific capture IgM and IgG ELISA in the differentiation of primary and secondary dengue virus infections showed good correlation (95.90% agreement). Most important, we have found that the serotype of the dengue virus from the majority of patients with primary infection could be correctly identified when convalescent-phase or postinfection sera were analyzed by NS1 serotype-specific IgG ELISA. These findings suggested that NS1 serotype-specific IgG ELISA could be reliably applied for serodiagnosis and seroepidemiological study of dengue virus infection.

Sensitive microplate enzyme-linked immunosorbent assay for detection of antibodies against the scrub typhus rickettsia, Rickettsia tsutsugamushi

1979 ◽  
Vol 9 (1) ◽  
pp. 38-48 ◽  
Author(s):  
G A Dasch ◽  
S Halle ◽  
A L Bourgeois
Keyword(s):  
Optical Density ◽  
Immunosorbent Assay ◽  
Scrub Typhus ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Serum Dilution ◽  
End Point

A microtiter enzyme-linked immunosorbent assay (ELISA) has been developed for the titration of antibodies against scrub typhus in human and animal sera. Scrub typhus rickettsiae were grown in monolayers of irradiated mouse LM3 cells and separated from host cell materials by differential centrifugation, filtration through a glass filter (AP-20, Millipore Corp.), and isopycnic banding in Renografin density gradients. The scrub typhus ELISA antigens were obtained from the purified viable rickettsiae by French pressure cell disruption and addition of 0.2% Formalin to the soluble extract. Antisera prepared in rabbits against the prototype Karp, the Kato, and the Gilliam strains of scrub typhus were used to standardize the ELISA and to compare its sensitivity and specificity to that of the indirect fluorescent antibody test (IFA). ELISA titers were measured as the greatest serum dilution showing an optical density 0.25 above controls or by the optical density achieved at a fixed serum dilution. The IFA and ELISA end point titers were quite similar, and all three measures of titer had comparable specificity for the strains of scrub typhus. No cross-reactions between the typhus and scrub typhus wera were observed by ELISA. Both the immunoglobulin M (IgM) and IgG antibody titers of 12 sequential sera from four patients with scrub typhus were obtained by IFA and ELISA. The IFA and ELISA end point titers for IgM and IgG had correlation coefficients of 0.91 and 0.97, respectively, whereas the ELISA optical density values at a serum dilution of 1:100 had slightly lower correlations with IFA titers (0.80 and 0.94). Early rising IgM titers followed by rising IgG titers were demonstrated by ELISA in three patients with primary scrub typhus infections, whereas the IgG response predominated in a patient with a reinfection. It is concluded that the ELISA for scrub typhus is a very satisfactory alternative to the IFA test.

scholarly journals Development of a Human-Murine Chimeric Immunoglobulin M for Use in the Serological Detection of Human Alphavirus Antibodies

2011 ◽  
Vol 18 (12) ◽  
pp. 2181-2182 ◽  
Author(s):  
Brett A. Thibodeaux ◽  
Nathan M. Liss ◽  
Amanda N. Panella ◽  
John T. Roehrig
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Immunoglobulin M ◽  
Murine Monoclonal Antibody ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Detection ◽  
Antibody Capture

ABSTRACTDiagnosis of human alphaviral infections relies on serological techniques, such as the immunoglobulin M antibody capture–enzyme-linked immunosorbent assay (MAC-ELISA). We have humanized the alphavirus broadly cross-reactive murine monoclonal antibody 1A4B-6 to create a reagent capable of replacing human positive sera in the MAC-ELISA for diagnosis of human alphaviral infections.

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