scholarly journals The Evolution of the Enzyme Immunoassay/Enzyme-Linked Immunosorbent Assay

2021 ◽  
Vol 2 (3) ◽  
pp. 13-17
Author(s):  
Leonid Tarassishin
Keyword(s):  
Enzyme Immunoassay ◽  
Immunosorbent Assay ◽  
High Specificity ◽  
Short Review ◽  
Immunological Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Target Antigen ◽  
Simple Method ◽  
Antibody Complex ◽  
Low Sensitivity

50 years ago the Enzyme Immunoassay Enzyme-Linked Immunosorbent Assay, mostly known as ELISA was developed. This is a powerful but simple method that is very widely used in the diagnostic practice, as well as in biomedical research. During this time a number of ELISA modification were developed that significantly increased its properties, especially the senstivity, such as avidin-biotin assay, immuno-PCR, nano-ELISA and finally, the digital ELISA. This short review describes the principles of ELISA and the evolution from a conventional assay to the modern ultra-sensitive method. Most of the immunological methods have two components: antigen and antibody. The high specificity of their interaction gives a possibility to detect one of them if other one is included in the reaction as a specific partner. The simplest method for antigen detection in the presence of the antibody is immune diffusion (radial immune diffusion in that case), which practically the formation of precipitate of the “antigen-antibody” complex, when the target antigen diffuses from well into agarose containing the specific antibody. Unfortunately, this assay, as well as other traditional methods, like hemagglutination or complement fixation, have a low sensitivity and are unwieldy.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay Method for the Detection of Rhein in Rheum officinale

2020 ◽  
Vol 2020 ◽  
pp. 1-7
Author(s):  
Min Chen ◽  
Tie-Gui Nan ◽  
Jie Xin ◽  
Li Cui ◽  
Bo Zhang ◽  
...  
Keyword(s):  
Quality Control ◽  
Immunosorbent Assay ◽  
High Specificity ◽  
Cross Reactivity ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Simple Method ◽  
Specificity And Sensitivity ◽  
Important Quality ◽  
Aloe Emodin

Rhein is an important quality-control marker of Rheum officinale. The aim of this study was to develop an indirect competitive enzyme-linked immunosorbent assay (icELISA) for rhein detection, which acts as a powerful tool for quality control and proper usage of Rheum officinale. First, a specific and sensitive monoclonal antibody (mAb) against rhein was produced from a stable hybridoma cell line, 1F8, generated by the fusion of mouse myeloma sp2/0 with spleen cells obtained from a Bal b/c mouse immunized with rhein-BSA. Then, an icELISA method was developed with an IC50 value and working range of 0.05 μg L−1 and 0.02–0.11 μg L−1, respectively. The icELISA revealed high assay specificity, since it only had a relatively high cross reactivity with aloe-emodin (27%) and almost no cross reactivity with any other anthraquinones (<1%). When spiked with 0.2–2 mg kg−1 of rhein, the recoveries ranged from 84.19% to 102.90%. Finally, icELISA was used to detect rhein contents of Rheum officinale collected from different regions, and the results corresponded well with those of HPLC. Overall, the developed icELISA with high specificity and sensitivity provided a rapid and simple method for rhein detection, and it may be a powerful tool for quality control and proper usage of Rheum officinale.

scholarly journals Analysis of the effectiveness of the use of enzyme-linked immunosorbent assay for the diagnosis of leukemia in cattle during health-improving activities

2020 ◽  
pp. 95-102
Author(s):  
S. I. Loginov
Keyword(s):  
Enzyme Immunoassay ◽  
Immunosorbent Assay ◽  
Serological Test ◽  
Leukemia Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Agricultural Enterprises ◽  
Low Level ◽  
Speed Up ◽  
Disease Indicators ◽  
Low Sensitivity

The aim of the work is to analyze the efficiency of using enzyme-linked immunosorbent assay in the diagnosis of cattle leukemia when carrying out health-improving measures in livestock enterprises that are unfavorable for this disease. Indicators of infection of cattle with leukemia virus on 6 farms of agricultural enterprises of the Tomsk region are presented. For serological diagnostics, the immunodiffusion reaction and enzyme immunoassay were used. It has been established that while carrying out health-improving measures in enterprises unfavorable for leukemia in cattle, the use of enzyme-linked immunosorbent assay for serological diagnosis of the disease enabled to identify a greater number of seropositive animals in comparison with a low-sensitivity immunodiffusion reaction. With a decrease in the herd infection rate to single cases of detection of animals infected with the leukemia virus at the final stages of rehabilitation of the enterprise, the number of additional seropositive animals detected by enzyme immunoassay increases. In the final period of herd recovery, the number of animals with a low level of antibodies to the leukemia virus, inaccessible to detection by immunodiffusion, increases. The use of an expensive and labor-consuming delivery of an enzymelinked immunosorbent assay to detect antibodies to the cattle leukemia virus is advisable at the final stages of an enterprise’s recovery. This enables to identify animals with a low level of antibodies to the leukemia virus, to speed up the negative result of a serological test of the entire herd and to exclude repeated outbreaks of infection of animals with this virus in a rehabilitated enterprise.

Multicolor enzyme-linked immunoassay method for visual readout of carbendazim

2021 ◽  
Author(s):  
Haoran Liu ◽  
Yiwen Wang ◽  
Ruijie Fu ◽  
Jing Zhou ◽  
Yanlin Liu ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity

Enzyme-linked immunosorbent assay (ELISA) with high specificity and sensitivity is one of the most popular techniques for detecting carbendazim (CBD), a commonly used benzimidazole fungicide in agriculture. However, the traditional...

scholarly journals Interpretations of Antibody Responses toSalmonella enterica Serotype Enteritidis gm Flagellin in Poultry Flocks Are Enhanced by a Kinetics-Based Enzyme-Linked Immunosorbent Assay

1998 ◽  
Vol 5 (4) ◽  
pp. 550-555 ◽  
Author(s):  
Patrick L. McDonough ◽  
Richard H. Jacobson ◽  
John F. Timoney ◽  
Ahmed Mutalib ◽  
David C. Kradel ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
High Sensitivity ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Department Of Agriculture ◽  
Trace Back ◽  
Elisa Format ◽  
Commercial Poultry ◽  
Computer Controlled

ABSTRACT Many regulatory and diagnostic programs for the detection ofSalmonella enterica serotype Enteritidis infection in commercial poultry flocks have relied on rapid Pullorum agglutination tests to screen birds because of the shared antigens of S. enterica Enteritidis and S. enterica Pullorum and Gallinarum; however, the use of the enzyme-linked immunosorbent assay (ELISA) format affords better analytical sensitivity than crude agglutination tests. In this study, we adapted our earlier conventional indirect ELISA, using gm flagellin as the antigen, to a kinetics-based, computer-controlled ELISA (KELA). The KELA was used to screen for flagellin antibody from three commercial flocks: (i) a large flock involved in a U.S. Department of Agriculture trace back from a humanS. enterica Enteritidis foodborne outbreak (n = 3,209), (ii) a flock infected with the endemicS. enterica Enteritidis serotype but which also had multiple other salmonella serotypes (n = 65), and (iii) an S. enterica Pullorum-infected flock (n = 12). The first flock (S. entericaEnteritidis prevalence of 2.45% based on culture) provided a field test of the KELA and allowed the calculation of diagnostic sensitivity (D-Sn) and diagnostic specificity (D-Sp). With a cutoff of 10 (used for screening flocks [i.e., high sensitivity]), the KELA has a D-Sn of 95.2% and a D-Sp of 18.5%; with a cutoff of 140 (used in confirmatory flock testing [i.e., high specificity]), the KELA has a D-Sn of 28.0% and a D-Sp of 99.1%. We found that with a cutoff of 60 (D-Sn = 63.1%; D-Sp = 91.6%), we could eliminate reactions in the KELA caused by other non-S. enterica Enteritidis salmonellae. The KELA was also compared to two commercial rapid Pullorum tests, the Solvay (D-Sn = 94.9%; D-Sp = 55.5%) and the Vineland (D-Sn = 62.0%; D-Sp = 75.3%).

scholarly journals Enzyme linked immunosorbent assay for rubella antibodies: a simple method of antigen production. A preliminary report

1995 ◽  
Vol 37 (4) ◽  
pp. 357-359 ◽  
Author(s):  
Vanda Akico Ueda Fick de Souza ◽  
Laura Masami Sumita ◽  
Mary Eiko S. Otsubo ◽  
Kioko Takei ◽  
Cláudio Sérgio Pannuti
Keyword(s):  
Enzyme Immunoassay ◽  
Correlation Coefficient ◽  
Filter Paper ◽  
Preliminary Report ◽  
Enzyme Linked Immunosorbent Assay ◽  
Simple Method ◽  
Blood Samples ◽  
Commercial Test ◽  
Antigen Production ◽  
Commercial Kit

A simple method of rubella antigen production by treatment with sodium desoxycholate for use in enzyme immunoassay (IMT-ELISA) is presented. When this assay was compared with a commercial test (Enzygnost-Rubella, Behring), in the study of 108 sera and 118 filter paper blood samples, 96.9% (219/226) overall agreement and correlation coefficient of 0.90 between absorbances were observed. Seven samples showed discordant results, negative by the commercial kit and positive by our test. Four of those 7 samples were available, being 3 positive by HI.

scholarly journals Comparison of an Enzyme Immunoassay versus Mass Spectrometry-based Assay for the Quantitative Determination of the Procollagen Type I N-terminal Propeptide in Rat Serum

2009 ◽  
Vol 2 ◽  
pp. PRI.S3454 ◽  
Author(s):  
Monika Dzieciatkowska ◽  
Marci Copeland ◽  
Jinsam You ◽  
Jean-Pierre Wery ◽  
Mu Wang
Keyword(s):  
Mass Spectrometry ◽  
Enzyme Immunoassay ◽  
Dynamic Range ◽  
High Specificity ◽  
Specific Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Type I ◽  
Rat Serum ◽  
Procollagen Type ◽  
Procollagen Type I

Traditionally, antibody-based assays, such as enzyme immunoassay (EIA), enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA), are the primary tool for the targeted quantification of a specific protein. An antibody-based assay can be run at high-throughput and has extraordinary sensitivity and specificity. In the cases where antibody-based assays exist, the process of validating biomarker candidates can be relatively straightforward. However, the antibody-based approach is limited by the lack of availability of antibodies with high specificity. The development of a high quality antibody-based assays can be costly, time-consuming and a resource-intensive effort. Another disadvantage of antibody-based assays is that they often do not discriminate closely related isoforms. While the antibody development is central to the success of antibody-based platform, mass spectrometry (MS) provides alternative and complementary approach to existing antibody-based assays. The MS-based assays are becoming very popular for quantitative candidates proteins detection in a complex biological mixture. In the present paper, an in-house developed mass spectrometry (MS)-based assay was compared to a commercially available EIA in reproducibility, measurement accuracy, and dynamic range using rat procollagen type-I N-terminal propeptide (P1NP) as a model.

scholarly journals Fluorescent Immunoassays for Detection and Quantification of Cardiac Troponin I: A Short Review

Molecules ◽  
2021 ◽  
Vol 26 (16) ◽  
pp. 4812
Author(s):  
Remya Radha ◽  
Syeda Kiran Shahzadi ◽  
Mohammad Hussein Al-Sayah
Keyword(s):  
Cardiovascular Diseases ◽  
Cardiac Troponin ◽  
Troponin I ◽  
Cardiac Injury ◽  
High Specificity ◽  
Short Review ◽  
Cardiac Troponin I ◽  
Diagnostic Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Assays

Cardiovascular diseases are considered one of the major causes of human death globally. Myocardial infarction (MI), characterized by a diminished flow of blood to the heart, presents the highest rate of morbidity and mortality among all other cardiovascular diseases. These fatal effects have triggered the need for early diagnosis of appropriate biomarkers so that countermeasures can be taken. Cardiac troponin, the central key element of muscle regulation and contraction, is the most specific biomarker for cardiac injury and is considered the “gold standard”. Due to its high specificity, the measurement of cardiac troponin levels has become the predominant indicator of MI. Various forms of diagnostic methods have been developed so far, including chemiluminescence, fluorescence immunoassay, enzyme-linked immunosorbent assay, surface plasmon resonance, electrical detection, and colorimetric protein assays. However, fluorescence-based immunoassays are considered fast, accurate and most sensitive of all in the determination of cardiac troponins post-MI. This review represents the strategies, methods and levels of detection involved in the reported fluorescence-based immunoassays for the detection of cardiac troponin I.

scholarly journals Enzyme Linked Immunosorbent Assay (ELISA) Technique Guideline

2021 ◽  
Vol 5 (5) ◽  
pp. 447-453
Author(s):  
Rachmat Hidayat ◽  
Patricia Wulandari
Keyword(s):  
Immunosorbent Assay ◽  
Solid Surface ◽  
Color Change ◽  
Antibody Binding ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hormone Levels ◽  
Elisa Technique ◽  
Antibody Complex ◽  
Antigen Antibody ◽  
Detection Signal

ELISA (Enzyme-linked immunosorbent assay) is a technique used to assess the quantification of peptide, protein, antibody and hormone levels, based on the principle of antigen-antibody binding. In the ELISA technique, antigen immobilization will be carried out on a solid surface, then bound with antibodies to form an antigen-antibody bond complex, where the antigen-antibody complex is bound to the enzyme. The detection signal in the form of a color change will be formed due to the reaction between the enzyme and the substrate.

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