scholarly journals Combined Cell Culture Enzyme-Linked Immunosorbent Assay for Quantification of Poliovirus Neutralization- Relevant Antibodies

2000 ◽  
Vol 7 (6) ◽  
pp. 915-919 ◽  
Author(s):  
A. F. Wahby
Keyword(s):  
Cell Culture ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Humoral Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Titration Curves ◽  
Microneutralization Assay ◽  
Human Sera ◽  
Reference Serum ◽  
Presence And Absence

ABSTRACT A combined cell culture enzyme-linked immunosorbent assay (CCC-ELISA) was developed for measuring the neutralizing antipoliovirus antibodies in human sera. The binding of different concentrations of each of the three poliovirus types to BGM cells in the presence and absence of a constant dilution from each test and reference serum was measured in the CCC-ELISA. The titers of the viruses neutralized by each serum were measured with the titration curves and used for interpretation of neutralizing titers to the three poliovirus types. Analysis of human sera revealed that the sensitivity and specificity of the CCC-ELISA and the microneutralization assay were comparable. The CCC-ELISA is nonsubjective, rapid, and highly reproducible. Furthermore, the CCC-ELISA could potentially be used as a seroepidemiologic tool for assessment of the humoral response to the cell culture infectious viruses.

scholarly journals Sensitive Enzyme-Linked Immunosorbent Assay for Detection of Antibodies Against Typhus Rickettsiae, Rickettsia prowazekii and Rickettsia typhi

1977 ◽  
Vol 6 (2) ◽  
pp. 101-110
Author(s):  
Sidney Halle ◽  
Gregory A. Dasch ◽  
Emilio Weiss
Keyword(s):  
Immunosorbent Assay ◽  
Complement Fixation ◽  
Differential Susceptibility ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rickettsia Prowazekii ◽  
Rickettsia Typhi ◽  
Titration Curves ◽  
Ether Extraction ◽  
Human Sera

An enzyme-linked immunosorbent assay (ELISA) has been developed for the titration of rickettsial antibodies in human and animal sera. Two preparations of soluble typhus-group antigens were obtained from Rickettsia typhi and Rickettsia prowazekii by ether extraction: a standard antigen from infected yolk sacs (YS antigen) and one free of yolk sac contaminants from Renografin-purified rickettsiae (PR antigen). Rabbit, mouse, and guinea pig sera were obtained by immunization with viable purified R. typhi or R. prowazekii . Human sera were obtained from individuals who had recovered from laboratory infections with either typhus rickettsia months or years previously. Goat-derived anti-immunoglobulins were conjugated to alkaline phosphatase with glutaraldehyde. Although the PR and YS antigens gave equivalent antibody titers in the complement fixation test, the PR antigen was clearly superior in the ELISA. With this antigen, the titration curves of all antisera were linear over a wider range of serum concentrations and the titers were higher than with the YS antigen. With YS and PR antigens, ELISA titers were higher than those obtained by complement fixation by one and two orders of magnitude, respectively. In human sera, immunoglobulin G and immunoglobulin M antibodies were demonstrated by their respective anti-immunoglobulins and by differential susceptibility to ethanethiol. ELISA titers showed some type specificity, whereas none was observed in complement fixation tests. The ELISA is highly sensitive, reproducible, and easily adaptable to the various requirements of clinical and research laboratories.

Detection of Toscana Virus-Specific Immunoglobulins G and M by an Enzyme-Linked Immunosorbent Assay Based on Recombinant Viral Nucleoprotein

1999 ◽  
Vol 37 (6) ◽  
pp. 2010-2012 ◽  
Author(s):  
Maria Grazia Ciufolini ◽  
Cristiano Fiorentini ◽  
Paola di Bonito ◽  
Stefania Mochi ◽  
Colomba Giorgi
Keyword(s):  
Infected Mouse ◽  
Sensitivity And Specificity ◽  
Mouse Brain ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Toscana Virus ◽  
Human Sera

An enzyme-linked immunosorbent assay (ELISA) based on the recombinant Toscana virus nucleoprotein (rN) has been developed. Its sensitivity and specificity for the detection of virus-specific immunoglobulins G and M in human sera were similar to those of the ELISA that is based on an antigen extracted from infected mouse brain and that is routinely used for serodiagnosis.

scholarly journals Patients with Pulmonary Tuberculosis Develop a Strong Humoral Response against Methylated Heparin-Binding Hemagglutinin

2005 ◽  
Vol 12 (9) ◽  
pp. 1135-1138 ◽  
Author(s):  
Stefania Zanetti ◽  
Alessandra Bua ◽  
Giovanni Delogu ◽  
Cinzia Pusceddu ◽  
Maristella Mura ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Immunosorbent Assay ◽  
Humoral Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Heparin Binding ◽  
Recombinant Antigens ◽  
Human Sera

ABSTRACT Reactivities of human sera against selected recombinant Mycobacterium tuberculosis antigens were assessed by enzyme-linked immunosorbent assay. The results obtained indicate that patients with tuberculosis (TB) do not develop a strong humoral response against PE_PGRS and PPE proteins or against the Ag85B and heparin-binding hemagglutinin (HBHA) recombinant antigens. Conversely, purified methylated HBHA was strongly recognized by sera obtained from TB patients compared to controls.

Integration of Liquid Chromatography with Immunoassay: An Approach Combining the Strengths of Both Methods

1994 ◽  
Vol 77 (5) ◽  
pp. 1275-1287 ◽  
Author(s):  
Petra M Krämer ◽  
Qing X Li ◽  
Bruce D Hammock
Keyword(s):  
Liquid Chromatography ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Uv Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Multiresidue Analysis ◽  
Selective Method ◽  
Separation Power ◽  
Immunochemical Detection ◽  
Very High

Abstract The integration of liquid chromatography (LC) with immunochemical detection combines the superior separation power of LC and the sensitivity and specificity of immunoassays. This approach is shown with 3 LC systems (Perkin-Elmer, C18 RP, 4.6 mm; Varian, C18 RP, 1 mm microbore; Michrom, C18 RP, 1 mm microbore) Integrated with an enzyme-linked immunosorbent assay (ELISA) selective for five 4-nitrophenols. The nitrophenols were separated with the 3 LC systems with isocratic runs of 15 to 20 min. Microbore LC separation showed a 10-20 times reduction in solvent amount compared to conventional separation. LC–immunoassay was about 8- to 10-fold more sensitive compared with LC with UV detection. Integrated LC–immunoassay proved to be a very selective method when 2-methylphenol was injected with an equimolar mixture of 2-amino-4-nitrophenol and 3-methyl-4-nrtrophenol; 2-methy I phenol does not crossreact with the serum used. Only 2 peaks could be seen in the detection, even when 2-methylphenol was present in very high amounts (3000 pmol). Further, the EUSA-LC detection proved to be selective and sensitive for complex matrixes. 2-Amlno-4-nitrophenol was clearly identified in spiked extracts of soil and plant, even when a very small amount (2.4 ng) was injected. Although LC–immunoassay is more labor intensive than LC with UV detection, it offers great advantages in multiresidue analysis and is generally applicable for peak confirmation.

scholarly journals Effect of Antigen Coating Conditions on Enzyme-Linked Immunosorbent Assay for Detection of Immunoglobulin G Antibody to Neisseria meningitidis Serogroup Y and W135 Capsular Polysaccharide Antigens in Serum

2003 ◽  
Vol 10 (6) ◽  
pp. 1136-1140 ◽  
Author(s):  
Peter C. Giardina ◽  
Renee E. Evans ◽  
Daniel J. Sikkema ◽  
Dace Madore ◽  
Stephen W. Hildreth
Keyword(s):  
Immunoglobulin G ◽  
Capsular Polysaccharide ◽  
Enzyme Linked Immunosorbent Assay ◽  
Meningococcal Vaccine ◽  
Igg Antibody ◽  
Human Sera ◽  
Polysaccharide Antigens ◽  
Reference Serum ◽  
Adult Volunteers ◽  
Assay Conditions

ABSTRACT Human sera collected from 28 consenting adult volunteers were used to define assay conditions for meningococcal vaccine clinical trial serology. Immunoassay parameters were optimized with these test sera and the standard reference serum, CDC1992. Coating conditions for serogroup Y and W135 polysaccharide antigens were found to influence the predicted serum immunoglobulin G (IgG) antibody concentrations. Sera that displayed IgG antibody binding profiles most unlike that of CDC1992 were influenced the most by coating conditions. Our results suggest that presentation of specific epitopes is influenced by antigen-coating concentrations for serogroup Y and W135 polysaccharides.

scholarly journals Assignment of Additional Anticapsular Antibody Concentrations to the Neisseria meningitidis Group A, C, Y, and W-135 Meningococcal Standard Reference Serum CDC1992

2002 ◽  
Vol 9 (3) ◽  
pp. 725-726 ◽  
Author(s):  
Cheryl M. Elie ◽  
Patricia K. Holder ◽  
Sandra Romero-Steiner ◽  
George M. Carlone
Keyword(s):  
Quality Control ◽  
Disease Control ◽  
Neisseria Meningitidis ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control And Prevention ◽  
Group A ◽  
Reference Serum ◽  
Centers For Disease Control

ABSTRACT We assigned additional enzyme-linked immunosorbent assay antibody concentrations (immunoglobulin G [IgG], IgM, and IgA, and total) to the Neisseria meningitidis standard reference serum CDC1992 for groups Y and W-135 to 12 Centers for Disease Control and Prevention quality control sera. These assignments will supplement previous assignments and will aid in the evaluation of present and developing vaccines.

scholarly journals Analysis ofLeishmaniamimetic neoglycoproteins for the cutaneous leishmaniasis diagnosis

Parasitology ◽  
2018 ◽  
Vol 145 (14) ◽  
pp. 1938-1948 ◽  
Author(s):  
Lígia Moraes Barizon de Souza ◽  
Vanete Thomaz Soccol ◽  
Ricardo Rasmussen Petterle ◽  
Michelle D. Bates ◽  
Paul A. Bates
Keyword(s):  
Cell Surface ◽  
Cutaneous Leishmaniasis ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Serum Levels ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
Cell Surfaces ◽  
Cell Markers ◽  
Antibody Titres

AbstractOligosaccharides are broadly present onLeishmaniacell surfaces. They can be useful for the leishmaniases diagnosis and also helpful in identifying new cell markers for the disease. The disaccharide Galα1-3Galβis the immunodominant saccharide inLeishmaniacell surface and is the unique non-reducing terminal glycosphingolipids structure recognized by anti-α-Gal. This study describes an enzyme-linked immunosorbent assay (ELISA) used to measure serum levels of anti-α-galactosyl (α-Gal) antibodies in patients with cutaneous leishmaniasis (CL). Optimal ELISA conditions were established and two neoglycoproteins (NGP) containing the Galα1-3Gal terminal fraction (Galα1-3Galβ1-4GlcNAc-HAS and Galα1-3Gal-HAS) and one Galα1-3Gal NGP analogue (Galα1-3Galβ1-3GlcNAc-HAS) were used as antigens. Means of anti-α-Gal antibody titres of CL patients were significantly higher (P< 0.05) than the healthy individuals for all NGPs tested. Sensitivity and specificity of all NGPs ranged from 62.2 to 78.4% and 58.3 to 96.7%, respectively. In conclusion, the NGPs can be used for CL diagnosis.

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