Development of an enzyme-linked immunosorbent assay for the detection of IgG-antibodies to the causative agent of COVID-19 in human serum (plasma)

2020 ◽  
Vol 65 (11) ◽  
pp. 683-687
Author(s):  
S. G. Mardanly ◽  
A. S. Avdonina ◽  
S. G. Mamedova
Keyword(s):  
Human Serum ◽  
Immunosorbent Assay ◽  
Causative Agent ◽  
Solid Phase ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Efficiency ◽  
Igg Antibodies ◽  
Comparative Tests ◽  
Test Kit

A new original Russian test kit for the detection of IgG-antibodies to the causative agent of COVID-19 - coronavirus SARS-CoV-2 by the method of enzyme-linked immunosorbent assay (ELISA) on a solid-phase «ELISA-SARS-CoV-2-AT-G» has been developed. In comparative tests with similar test systems «Vitrotest® SARS-CoV-2 IgG» (Vitrotest, Ukraine) and «Anti-SARS-Cov-2 ELISA (IgG)» (EUROIMMUN AG, Germany) high diagnostic efficiency of the new test system was shown.

scholarly journals Development of the ELISA Test System for Quantitative Determination of IgG to the Varizella zoster virus in Human Serum and Assessment of its Diagnostic Efficiency

2022 ◽  
Vol 20 (6) ◽  
pp. 72-80
Author(s):  
L. N. Lukhverchyk ◽  
G. L. Alatortseva ◽  
L. N. Nesterenko ◽  
V. Y. Kabargina ◽  
V. V. Dotsenko ◽  
...  
Keyword(s):  
Herpes Zoster ◽  
Blood Serum ◽  
Human Serum ◽  
Immunosorbent Assay ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Efficiency ◽  
Functional Characteristics ◽  
Serum Samples ◽  
Population Immunity

Relevance. The introduction of Varicella vaccine prophylaxis explains the need to develop a methodology for monitoring the vaccination effectiveness and the intensity of population immunity. This problem can be solved using quantitative immunoassay methods. Aim. Development of an enzyme-linked immunosorbent assay for the concentration of class G immunoglobulins (AB) to Varicella zoster virus (VZV) determining and assessing its functional characteristics and diagnostic efficiency. Materials and methods. Recombinant antigen GE VZV. WHO International Standard for Antibodies to VZV W1044. Blood serum samples from healthy people and patients with Chickenpox and Herpes zoster, blood serum samples containing IgG antibodies to herpes simplex viruses of the first and second types, cytomegalovirus, Epstein-Barr virus. Anti-VZV ELISA (IgG) reagent kit (Euroimmun, Germany). Indirect enzyme-linked immunosorbent assay. Immunization of animals with recombinant antigen GE, isolation, and purification of specific antibodies. Conjugation of monoclonal antibodies to human IgG with antibodies to antigen GE and with horseradish peroxidase. Results. An enzyme-linked immunosorbent assay in «an indirect» format has been developed to determine the specific antibodies to VZV concentration (IU/ml) in human serum/plasma. An artificial calibrator for determining the concentration of AB-VZV had been synthesized and standardized according to the International WHO-standard W1044. The main functional characteristics of the developed enzyme-linked immunosorbent assay are determined in accordance with GOST 51352-2013. The diagnostic kit was tested on blood serum samples from children with chickenpox (n = 43), adults with Herpes zoster (n = 158), healthy individuals (n = 781). The diagnostic sensitivity of the test system was 85%, the diagnostic specificity was 87% according to the ROC analysis. The absence of cross-reactivity of the test system was shown on samples with serological markers of other herpesvirus infections (n = 94). Comparative trials of the developed test system and its commercial analog, the Anti-VZV ELISA (IgG) reagent kit, did not reveal statistically significant differences between their functional characteristics. Conclusions. The developed test system for determining of the AB-VZV concentration in human serum/plasma in terms of its functional characteristics meets the GOST requirements, is characterized by high diagnostic efficiency, can be used to monitor the effectiveness of vaccine prophylaxis and strength of population immunity, as well as to assess the immune response in chickenpox and Herpes zoster.

scholarly journals Line immunoassay for detection of IgG antibodies to hepatitis E virus (Hepeviridae, Orthohepevirus, Orthohepevirus A)

2020 ◽  
Vol 65 (3) ◽  
pp. 132-142
Author(s):  
G. I. Alatortseva ◽  
V. V. Dotsenko ◽  
L. N. Nesterenko ◽  
L. N. Luhverchik ◽  
V. Yu. Kabargina ◽  
...  
Keyword(s):  
Human Serum ◽  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Test System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Igg Antibodies ◽  
Recombinant Antigens ◽  
Test Kit ◽  
Wide Range

Introduction. The diagnostic efficacy of methods for hepatitis E serodiagnostic varies over a wide range; therefore, the combined use of tests of various formats is recommended. The aim of the research was to develop a test system for the detection of IgG antibodies to hepatitis E virus (HEV) in human serum by linear immunoassay (LIA). Material and methods. Serum samples from patients with hepatitis and healthy individuals were tested using commercial enzyme-linked immunosorbent assay systems for the presence of IgG antibodies to viral agents causing hepatitis and other infections associated with liver pathology. Recombinant antigens ORF2 and ORF3 of HEV genotypes 1 and 3 were used. The “RecomLine HEV IgG/IgM” reagent kit (Mikrogen GmbH, Germany) was used as a comparison test system. Results. The first Russian diagnostic kit “Blot-HEV”, designed to detect IgG antibodies to individual HEV proteins in human serum using LIA, was developed. The antigenic base is represented by strips of a nitrocellulose membrane with immobilized recombinant antigens ORF2 (aa 406–660) and ORF3 (aa 1–113) of HEV genotypes 1 and 3, and control antigens in the form of discrete lines. The conjugate was mouse monoclonal antibodies to human class G immunoglobulins labeled with horseradish peroxidase. The chromogen solution contained the 3,3’,5,5’-tetramethylbenzidine. A visual and digital recording of results was provided. The analytical sensitivity of the test kit was 0.625 IU/ml for ORF2 antigens and 2.5 IU/ml for ORF3 antigens. The absence of the influence of endogenous interfering substances on the results of the analysis and the absence of cross-reactions with antibodies to hepatitis pathogens of the other etiologies had been shown. The sensitivity of the test system compared to the “RecomLine HEV IgG/IgM” kit was 92%, specificity 97%. Shelf life in condition of storage was determined to be 12 months. Conclusions. The developed test can be used to confirm the results of ELISA in laboratory diagnosis of hepatitis E.

scholarly journals Development and Validation of 13-plex Luminex-Based Assay for Measuring Human Serum Antibodies to Streptococcus pneumoniae Capsular Polysaccharides

mSphere ◽  
2018 ◽  
Vol 3 (4) ◽  
pp. e00128-18 ◽  
Author(s):  
Danka Pavliakova ◽  
Peter C. Giardina ◽  
Soraya Moghazeh ◽  
Shite Sebastian ◽  
Maya Koster ◽  
...  
Keyword(s):  
Human Serum ◽  
Lower Limit ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Correlate Of Protection ◽  
Immune Correlate ◽  
Relative Standard ◽  
Limit Of Quantitation

ABSTRACT A Luminex-based direct immunoassay (dLIA) platform has been developed to replace the standardized pneumococcal enzyme-linked immunosorbent assay platform. The multiplex dLIA simultaneously measures the concentration of serum immunoglobulin G (IgG) antibodies specific for pneumococcal capsular polysaccharide (PnPS) serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. The assay uses poly-l-lysine (PLL)-conjugated PnPS, chemically coupled to spectrally distinct Luminex microspheres. Assay validation experiments were performed using residual human serum samples obtained from 13-valent pneumococcal conjugate vaccine (13vPnC) clinical studies. Assay results are expressed as IgG antibody concentrations in micrograms per milliliter using the international reference serum, 007sp. The lower limit of quantitation (LLOQ) for all serotypes covered in the 13-plex dLIA fell within the range of 0.002 to 0.038 µg/ml serum IgG. The difference between the lower limit and upper limit of the assay range was >500-fold for all serotypes, and assay variability was <20% relative standard deviation (RSD) for all serotypes. IgG antibody measurements were shown to be serotype-specific (some cross-reactivity was observed only between the structurally related serotypes 6A and 6B as well as 19A and 19F), and no interference was observed between the serotypes when the assay was performed in the 13-plex format compared to the singleplex assays. The 13-plex dLIA platform developed by Pfizer Inc. generates up to 143 test results in a single 96-well plate and is a suitable replacement of the enzyme-linked immunosorbent assay (ELISA) platform for evaluating vaccine clinical trials. IMPORTANCE The pneumococcal enzyme-linked immunosorbent assay (ELISA) measures IgG antibodies in human serum, and it is an important assay that supports licensure of pneumococcal vaccines. The immune correlate of protection, 0.35 µg/ml of IgG antibodies, was determined by the ELISA method. Pfizer has developed a new Luminex-based assay platform to replace the ELISA. These papers describe the important work of (i) validating the Luminex-based assay and (ii) bridging the immune correlate of protection (0.35 µg/ml IgG) to equivalent values reported by the Luminex platform.

scholarly journals Detection of tetanus toxoid antibodies in human sera in New Zealand by ELISA

1987 ◽  
Vol 98 (2) ◽  
pp. 199-202 ◽  
Author(s):  
R. C. H. Lau
Keyword(s):  
New Zealand ◽  
Human Serum ◽  
Immunosorbent Assay ◽  
Tetanus Toxoid ◽  
Indirect Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Age Group ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Human Sera

SUMMARYAn enzyme-linked immunosorbent assay (ELISA) incorporating the sensitive biotin-streptavidin system was developed to detect IgG antibodies to tetanus toxoid in human serum. Serum samples obtained from 557 normal persons aged 1–65 years from different areas in New Zealand were tested. The proportion of those immune ranged from 60–93% in males, and from 46–86% in females. In the 1–9 years age group 85% were immune. The indirect ELISA is suitable for serological surveys as it is simple to perform, economical and reproducible.

A New Technique in Quantitative Immunohematology: Solid-Phase Kinetic Enzyme-Linked Immunosorbent Assay

Vox Sanguinis ◽  
1983 ◽  
Vol 45 (6) ◽  
pp. 440-448 ◽  
Author(s):  
S. Spitalnik ◽  
J. Cowles ◽  
M.T. Cox ◽  
D. Baker ◽  
J. Holt ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
New Technique ◽  
Enzyme Linked Immunosorbent Assay ◽  
A New Technique

Enzyme-linked immunosorbent assay for cholesteryl ester transfer protein in human serum

1995 ◽  
Vol 240 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Toshitaka Sato ◽  
Masayoshi Fukasawa ◽  
Makoto Kinoshita ◽  
Hiroyuki Arai ◽  
Takao Saeki ◽  
...  
Keyword(s):  
Cholesteryl Ester ◽  
Human Serum ◽  
Immunosorbent Assay ◽  
Cholesteryl Ester Transfer Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transfer Protein

Measles antibodies and response to vaccination in children aged less than 14 months: implications for age of vaccination

2011 ◽  
Vol 140 (9) ◽  
pp. 1599-1606 ◽  
Author(s):  
E. BORRÀS ◽  
L. URBIZTONDO ◽  
J. COSTA ◽  
J. BATALLA ◽  
N. TORNER ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Maternal Antibodies ◽  
Passive Immunity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Blood Samples ◽  
Measles Outbreak ◽  
Measles Antibodies ◽  
Measles Vaccination

SUMMARYPassive immunity against measles decreases during the first months of life. The objective of this study was to determine titres of measles antibodies in children aged 9–14 months and their mothers before vaccination, and the children's response to vaccination. Blood samples were collected by capillary puncture before and 28 days after vaccination. Samples were obtained between February and June 2007 during an ongoing measles outbreak. Titres of specific measles IgG antibodies were determined by enzyme-linked immunosorbent assay. Seroconversion was defined as the presence of antibodies after vaccination in subjects without antibodies before vaccination. Maternal antibodies were present in 37·7% of all 69 children included and in 45·1% of children aged 9 months. Of the 51 children in whom a second sample was obtained, 31 (60·8%) were seronegative before vaccination and 61·3% seroconverted. Interference of maternal antibodies was 30%. Advancing the first dose of measles vaccination from 15 to 12 months is a correct strategy, given the increase in the time of susceptibility of infants to measles.

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