Evaluation of the radioimmunoassay, indirect enzyme linked immunosorbent assay, and dot blot assay for the identification ofXanthomonas campestrispv.phaseoli

1985 ◽  
Vol 7 (3) ◽  
pp. 217-222 ◽  
Author(s):  
Eileen Malin ◽  
E. Lee Belden ◽  
Don Roth
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Dot Blot Assay

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay Using a Recombinant LigA Fragment Comprising Repeat Domains 4 to 7.5 as an Antigen for Diagnosis of Equine Leptospirosis

2013 ◽  
Vol 20 (8) ◽  
pp. 1143-1149 ◽  
Author(s):  
Weiwei Yan ◽  
Muhammad Hassan Saleem ◽  
Patrick McDonough ◽  
Sean P. McDonough ◽  
Thomas J. Divers ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Serum Dilution ◽  
Content Type ◽  
Diagnostic Antigen ◽  
Repeat Domain ◽  
Associated Proteins ◽  
Dot Blot Assay

ABSTRACTLeptospiraimmunoglobulin (Ig)-like (Lig) proteins are a novel family of surface-associated proteins in which the N-terminal 630 amino acids are conserved. In this study, we truncated the LigA conserved region into 7 fragments comprising the 1st to 3rd (LigACon1-3), 4th to 7.5th (LigACon4-7.5), 4th (LigACon4), 4.5th to 5.5th (LigACon4.5–5.5), 5.5th to 6.5th (LigACon5.5–6.5), 4th to 5th (LigACon4-5), and 6th to 7.5th (LigACon6-7.5) repeat domains. All 7 recombinant Lig proteins were screened using a slot-shaped dot blot assay for the diagnosis of equine leptospirosis. Our results showed that LigACon4-7.5 is the best candidate diagnostic antigen in a slot-shaped dot blot assay. LigACon4-7.5 was further evaluated as an indirect enzyme-linked immunosorbent assay (ELISA) antigen for the detection ofLeptospiraantibodies in equine sera. This assay was evaluated with equine sera (n= 60) that were microscopic agglutination test (MAT) negative and sera (n= 220) that were MAT positive to the 5 serovars that most commonly cause equine leptospirosis. The indirect ELISA results showed that at a single serum dilution of 1:250, the sensitivity and specificity of ELISA were 80.0% and 87.2%, respectively, compared to those of MAT. In conclusion, an indirect ELISA was developed utilizing a recombinant LigA fragment comprising the 4th to 7.5th repeat domain (LigACon4-7.5) as a diagnostic antigen for equine leptospirosis. This ELISA was found to be sensitive and specific, and it yielded results that concurred with those of the standard MAT.

Detection of M2 antimitochondrial antibodies by dot blot assay is more specific than by enzyme linked immunosorbent assay

2008 ◽  
Vol 56 (1) ◽  
pp. 10-14 ◽  
Author(s):  
I. Bargou ◽  
A. Mankaï ◽  
A. Jamaa ◽  
I. Ben Jazia ◽  
K. Skandrani ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antimitochondrial Antibodies ◽  
Dot Blot ◽  
Dot Blot Assay

A Simple, Rapid and Non-Radiolabeled Immune Assay to Detect Anti-AChR Antibodies in Myasthenia Gravis

2018 ◽  
Vol 50 (3) ◽  
pp. 229-235 ◽  
Author(s):  
Suresh Bokoliya ◽  
Shripad Patil ◽  
Madhu Nagappa ◽  
Arun Taly
Keyword(s):  
Myasthenia Gravis ◽  
Case Control Study ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
Dot Blot ◽  
Dot Blot Assay ◽  
Healthy Control ◽  
Neurological Illnesses ◽  
Control Study ◽  
Immune Assay

AbstractObjectiveTo assess the practicality of dot-blot testing for rapid and sensitive detection of the antiacetylcholine receptor (anti-AChR) antibodies in myasthenia gravis (MG).MethodsIn this case-control study, we tested serum specimens of 85 patients with MG, 85 healthy control individuals, and 85 patients without MG who have other autoimmune and neurological illnesses. All the serum specimens were tested for anti-AChR antibodies using 3 assays: in-house enzyme-linked immunosorbent assay (ELISA), the dot-blot assay, and commercial ELISA.ResultsIn-house ELISA, commercial ELISA, and dot-blot test results were positive for anti-AChR antibodies in 65 (76.5%) patients with MG. The results of all 3 tests were negative for anti-AChR antibodies in healthy controls and patients without MG. We observed perfect concordance (K = 1, P <.001) between all 3 tests. In-house ELISA correlated significantly (r = 0.873, P <.001) with commercial ELISA. In-house ELISA and the dot-blot test demonstrated similar diagnostic performance in detecting anti-AChR antibodies.ConclusionsThe dot-blot assay is a simple, nonradioactive immune assay for rapid detection of anti-AChR antibodies in MG.

scholarly journals Rapid Detection of Salmonella enterica Serovar Choleraesuis in Blood Cultures by a Dot Blot Enzyme-Linked Immunosorbent Assay

2000 ◽  
Vol 7 (6) ◽  
pp. 977-979 ◽  
Author(s):  
Kritsana Janyapoon ◽  
Sunee Korbsrisate ◽  
Hatairat Thamapa ◽  
Sittichai Thongmin ◽  
Suwattana Kanjanahareutai ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Salmonella Enterica ◽  
Rapid Detection ◽  
Direct Detection ◽  
Blood Cultures ◽  
Enzyme Linked Immunosorbent Assay ◽  
Biochemical Method ◽  
Dot Blot

ABSTRACT A dot blot enzyme-linked immunosorbent assay (ELISA) with a monoclonal antibody specific to phase1-c Salmonella was developed for the direct detection of Salmonella entericaserovar Choleraesuis in blood cultures. This system was applied to the identification of serovar Choleraesuis, and the results were compared with those obtained by a conventional biochemical method. It was revealed that all 12 samples identified to be infected with serovar Choleraesuis were positive on testing by the ELISA. In contrast, 77 samples infected with bacteria commonly isolated from the blood were not reactive by the ELISA. The calculated sensitivity and specificity of the established assay are 100%.

scholarly journals Generation and diagnostic application of monoclonal antibodies against Seneca Valley virus

2011 ◽  
Vol 24 (1) ◽  
pp. 42-50 ◽  
Author(s):  
Ming Yang ◽  
Rebekah van Bruggen ◽  
Wanhong Xu
Keyword(s):  
Monoclonal Antibodies ◽  
Indirect Elisa ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Specific Antibody Response ◽  
Dot Blot Assay ◽  
Infected Cells ◽  
Seneca Valley Virus ◽  
Vesicular Disease

Seneca Valley virus (SVV), a member of the Picornaviridae family, was implicated in a suspicious vesicular disease discovered in pigs from Canada in 2007. Because any outbreak of vesicular disease in pigs is assumed to be foot-and-mouth disease (FMD) until confirmed otherwise, a test for diagnosing the presence of SVV would be a very useful tool. To develop the diagnostic tests for SVV infection, 5 monoclonal antibodies (mAbs) were produced from mice immunized with binary ethylenimine (BEI)-inactivated SVV. Using a dot blot assay, the reactivity of the mAbs was confirmed to be specific for SVV, not reacting with any of the other vesicular disease viruses tested. The mAbs demonstrated reactivity with SVV antigen in infected cells by an immunohistochemistry assay. An SVV-specific competitive enzyme-linked immunosorbent assay (cELISA) was developed using BEI-inactivated SVV antigen and a mAb for serodiagnosis. The cELISA results were compared to the indirect isotype (immunoglobulin [Ig]M and IgG) ELISA and the virus neutralization test. All SVV experimentally inoculated pigs exhibited a positive SVV-specific antibody response at 6 days postinoculation, and the sera remained positive until the end of the experiment on day 57 (>40% inhibition) using the cELISA. The cELISA reflected the profile of the indirect ELISA for both IgM and IgG. This panel of SVV-specific mAbs is valuable for the identification of SVV antigen and the serological detection of SVV-specific antibodies.

scholarly journals Effect of Immunization with Recombinant OspA on Serologic Tests for Lyme Borreliosis

2001 ◽  
Vol 8 (1) ◽  
pp. 79-84 ◽  
Author(s):  
Paul T. Fawcett ◽  
Carlos D. Rose ◽  
Sandra M. Budd ◽  
Kathleen M. Gibney
Keyword(s):  
Western Blot ◽  
Lyme Borreliosis ◽  
Vaccine Recipient ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
Dot Blot ◽  
Western Blot Assay ◽  
Serologic Tests ◽  
Human Volunteers ◽  
Dot Blot Assay

ABSTRACT This study evaluated the effects of vaccination with OspA on the use of serologic tests as aids in the diagnosis of Lyme borreliosis. Sera from control and OspA-immunized mice and from OspA-immunized human volunteers were tested for serologic reactivity to Borrelia burgdorferi. Testing was performed with samples obtained prior to administration of vaccine and at 30 days following administration of an initial and a second dose of OspA vaccine. The assays used to assess serologic reactivity included an in-house-developed enzyme-linked immunosorbent assay (ELISA), an in-house-developed Western blot assay, two commercial Western blot tests, and a commercially available dot blot assay. Data obtained from this study demonstrate that immunization with the OspA vaccine will cause ELISA to yield positive results (as reported previously) for the majority of vaccine recipients. Results obtained from Western blot analysis indicate that vaccination with recombinant OspA induces production of antibodies which bind to several different borrelial proteins. The degree of reactivity detected by Western blotting varied greatly between the three assays used. The in-house assay showed the least reactivity, while one commercial Western blot test actually yielded positive test results for infection with B. burgdorferi. The usefulness of all three Western blot assays for the diagnosis of potential infection in a vaccine recipient is severely limited by the extensive reactivity caused by vaccination alone. Antibodies produced in response to OspA vaccination did not significantly affect the performance of the dot blot test; thus, it could provide a reliable means to test for infection withB. burgdorferi in OspA vaccine recipients.

scholarly journals Evaluation of Diagnostic Applications of Monoclonal Antibodies against Avian Influenza H7 Viruses

2010 ◽  
Vol 17 (9) ◽  
pp. 1398-1406 ◽  
Author(s):  
Ming Yang ◽  
Alfonso Clavijo ◽  
Jill Graham ◽  
John Pasick ◽  
James Neufeld ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Influenza A ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Western Blots ◽  
Positive Antibody ◽  
Dot Blot Assay ◽  
Diagnostic Applications ◽  
Ha Protein ◽  
H7 Subtype

ABSTRACT A panel of monoclonal antibodies (MAbs) was generated from mice immunized with binary ethylenimine (BEI)-inactivated H7N1 (A/TK/ON/18-2/00) virus. Using a dot blot assay, six of seven MAbs reacted with viruses of the H7 subtype, but not with any of the other 15 hemagglutinin (HA) subtypes tested. Four of the seven MAbs reacted with 14 different H7 isolates, indicating that the MAbs binding epitopes are conserved among viruses of the H7 subtype. The binding epitopes of all seven MAbs were conformational and reacted with the HA1 fraction of the HA protein in Western blots under nonreducing conditions. Applications of these MAbs in the development of rapid tests for H7 subtype viruses were evaluated. The MAbs demonstrated reactivity with AI virus H7 antigen in immunofluorescence and immunohistochemistry assays. Monoclonal antibody 3 showed a very strong immunostaining in the formalin-fixed and paraffin-embedded tissue from the H7N3 virus-infected chicken. A double-antibody sandwich (DAS) enzyme-linked immunosorbent assay (ELISA) was developed using two of the MAbs. The DAS ELISA specifically detected all H7 strains tested in this study. A competitive ELISA (cELISA) for the detection of H7-specific antibodies was evaluated using one MAb and BEI-inactivated H7N1 virus as the antigen. All infected birds showed positive antibody responses at 7 days postinfection. The sensitivity of this cELISA was comparable with that of an influenza A nucleoprotein-based cELISA. This panel of MAbs is valuable in the development of various immunoassays.

Development of an immunobinding dot-blot assay as an alternative for the serodiagnosis of human cysticercosis

2009 ◽  
Vol 83 (4) ◽  
pp. 333-337 ◽  
Author(s):  
E. Hernández-Cruz ◽  
J.J. González-Cabriales ◽  
C. Ordaz-Pichardo ◽  
N.I. de la Cruz-Hernández ◽  
G.H. Flores-Gutiérrez
Keyword(s):  
Toxoplasma Gondii ◽  
Gold Standard ◽  
Taenia Solium ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Validity ◽  
Dot Blot ◽  
Predictive Values ◽  
Cysticercus Cellulosae ◽  
Dot Blot Assay ◽  
Antigen Protein

AbstractAn immunobinding dot-blot assay (DBA) was developed on nitrocellulose paper for the serodiagnosis of human cysticercosis, using Cysticercus cellulosae as antigen. The DBA had an immunological sensitivity of 0.08 mg of antigen protein/ml; however, it showed cross-reactions with antigens of adult Taenia solium and Echinococcus granulosus, but not with Toxoplasma gondii and Entamoeba histolytica antigens. An enzyme-linked immunosorbent assay (ELISA) was used as the gold standard for obtaining the diagnostic validity of the DBA, giving 84.61%, 100.00%, 100.00% and 97.98% for epidemiological sensitivity, epidemiological specificity and positive and negative predictive values, respectively. There were no statistical differences between the two tests (P < 0.05, kappa = 0.907). This study showed that DBA is an alternative method for the serodiagnosis of human cysticercosis.

Sign in / Sign up

Export Citation Format

Share Document