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.

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 SARS-CoV-2 S1 and N-based serological assays reveal rapid seroconversion and induction of specific antibody response in COVID-19 patients

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Abdullah Algaissi ◽  
Mohamed A. Alfaleh ◽  
Sharif Hala ◽  
Turki S. Abujamel ◽  
Sawsan S. Alamri ◽  
...  
Keyword(s):  
Antibody Response ◽  
Specific Antibody ◽  
Large Scale ◽  
Disease Onset ◽  
High Specificity ◽  
Epidemiological Studies ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity ◽  
Human Coronaviruses

Abstract As the Coronavirus Disease 2019 (COVID-19), which is caused by the novel SARS-CoV-2, continues to spread rapidly around the world, there is a need for well validated serological assays that allow the detection of viral specific antibody responses in COVID-19 patients or recovered individuals. In this study, we established and used multiple indirect Enzyme Linked Immunosorbent Assay (ELISA)-based serological assays to study the antibody response in COVID-19 patients. In order to validate the assays we determined the cut off values, sensitivity and specificity of the assays using sera collected from pre-pandemic healthy controls, COVID-19 patients at different time points after disease-onset, and seropositive sera to other human coronaviruses (CoVs). The developed SARS-CoV-2 S1 subunit of the spike glycoprotein and nucleocapsid (N)-based ELISAs not only showed high specificity and sensitivity but also did not show any cross-reactivity with other CoVs. We also show that all RT-PCR confirmed COVID-19 patients tested in our study developed both virus specific IgM and IgG antibodies as early as week one after disease onset. Our data also suggest that the inclusion of both S1 and N in serological testing would capture as many potential SARS-CoV-2 positive cases as possible than using any of them alone. This is specifically important for tracing contacts and cases and conducting large-scale epidemiological studies to understand the true extent of virus spread in populations.

scholarly journals SARS-CoV-2 S1 and N-Based Serological Assays Reveal Rapid Seroconversion and Induction of Specific Antibody Response in COVID-19 Patients

2020 ◽  
Author(s):  
Abdullah Algaissi ◽  
Mohamed A. Alfaleh ◽  
Sherif Hala ◽  
Turki S. Abujamel ◽  
Sawsan S. Alamri ◽  
...  
Keyword(s):  
Antibody Response ◽  
Specific Antibody ◽  
Large Scale ◽  
Disease Onset ◽  
High Specificity ◽  
Epidemiological Studies ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity ◽  
Human Coronaviruses

As the coronavirus disease 2019 (COVID-19), which is caused by the novel SARS-CoV-2, continues to spread rapidly around the world, there is a need for well validated serological assays that allow the detection of viral specific antibody responses in COVID-19 patients or recovered individuals. In this study, we established and used multiple indirect Enzyme Linked Immunosorbent Assay (ELISA)-based serological assays to study the antibody response in COVID-19 patients. In order to validate the assays we determined the cut off values, sensitivity and specificity of the assays using sera collected from pre-pandemic healthy controls, COVID-19 patients at different time points after disease-onset, and seropositive sera to other human coronaviruses. The developed SARS-CoV-2 S1 subunit of the spike glycoprotein and nucleocapsid (N)-based ELISAs not only showed high specificity and sensitivity but also did not show any cross-reactivity with other CoVs. We also show that all RT-PCR confirmed COVID-19 patients tested in our study developed both virus specific IgM and IgG antibodies as early as week one after disease onset. Our data also suggest that the inclusion of both S1 and N in serological testing would capture as many potential SARS-CoV-2 positive cases as possible than using any of them alone. This is specifically important for tracing contacts and cases and conducting large-scale epidemiological studies to understand the true extent of virus spread in populations.

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 Multiple Indirect ELISAs for Serological Detection of SARS-CoV-2 Antibodies

2020 ◽  
Author(s):  
Abdullah Algaissi ◽  
Mohamed A. Alfaleh ◽  
Sherif Hala ◽  
Turki S. Abujamel ◽  
Sawsan S. Alamari ◽  
...  
Keyword(s):  
Antibody Response ◽  
Large Scale ◽  
Disease Onset ◽  
High Specificity ◽  
Epidemiological Studies ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Tests ◽  
Specificity And Sensitivity ◽  
Novel Coronavirus

As the coronavirus disease 2019 (COVID-19), which is caused by the novel coronavirus SARS-CoV-2, continues to spread rapidly around the world, there is an urgent need for validated serological assays to evaluate viral specific antibody responses in COVID-19 patients or recovered individuals. In this study, we established and used indirect Enzyme Linked Immunosorbent Assay (ELISA)-based serological tests to study the antibody response in COVID-19 patients. In order to validate the assays, we determined the cut-off values, sensitivity and specificity of the developed assays using sera collected from COVID-19 patients in Saudi Arabia at different time points after disease onset, as well as sera that are seropositive to other human CoVs; namely MERS-CoV, hCoV-OC43, hCoV-NL63, hCoV-229E, and hCoV-HKU1. The SARS-CoV-2 S1 subunit of the spike glycoprotein and nucleocapsid (N) ELISAs that we developed here not only showed high specificity and sensitivity, but also did not show any cross-reactivity with other CoVs. We also showed that all RT-PCR confirmed COVID-19 patients included in our study developed both virus specific IgM and IgG as early as one week after the onset of disease. The availability of these validated assays will enable us to determine the nature and duration of the antibody response mounted in response to SARS-CoV-2 infection. It will also allow conducting large-scale epidemiological studies to determine evidence of previous exposure to the virus and assess the true extent of virus spread within communities.

Development of an Enzyme-Linked Immunosorbent Assay Method To Detect Mustard Protein in Mustard Seed Oil

2007 ◽  
Vol 70 (1) ◽  
pp. 179-183 ◽  
Author(s):  
STEF J. KOPPELMAN ◽  
RIEK VLOOSWIJK ◽  
GINA BOTTGER ◽  
GERT van DUIJN ◽  
PETER van der SCHAFT ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Seed Oil ◽  
Food Product ◽  
Cross Reactivity ◽  
Polyclonal Antiserum ◽  
Assay Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mustard Seed ◽  
Calculated Level ◽  
Mustard Protein

An enzyme-linked immunosorbent assay for the detection of mustard protein was developed. The assay is based on a polyclonal antiserum directed against a mixture of mustard proteins raised in rabbits. The assay has a detection limit of 1.5 ppm (milligrams per kilogram) and is suitable for the detection of traces of mustard protein in mustard seed–derived flavoring ingredients. Limited cross-reactivity testing showed that no other plant proteins reacted significantly. From the animal proteins tested, only milk showed some cross-reactivity. With this sensitive assay, it was shown that refined mustard seed oil produced by steam distillation does not contain detectable amounts of mustard protein. Mustard seed oil is used as a flavoring in very low quantities, typically between 40 and 200 mg/kg. Thus, 100 g of a food product flavored with 200 mg of mustard seed oil per kg containing &lt;1.5 mg of protein per kg would represent an amount of mustard seed protein of &lt;30 ng. Taking into account the published literature on allergic reactions to the unintended ingestion of mustard, this conservatively low calculated level indicates that it is unlikely that food products containing mustard seed oil as a flavoring ingredient will elicit an allergic reaction in mustard-allergic individuals.

Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Detecting Tetrodotoxin

2012 ◽  
Vol 466-467 ◽  
pp. 220-224
Author(s):  
Qing Ping Zhong ◽  
Yan Ting Liu ◽  
Yuan Ming Sun ◽  
Hoi Fu Yu ◽  
Hong Tao Lei
Keyword(s):  
Monoclonal Antibody ◽  
Working Conditions ◽  
Immunosorbent Assay ◽  
Room Temperature ◽  
High Specificity ◽  
Keyhole Limpet Hemocyanin ◽  
Enzyme Linked Immunosorbent Assay ◽  
Coating Antigen ◽  
Specificity And Sensitivity ◽  
A Cell

This study aimed to develop a method for the detection of tetrodotoxin (TTX) based on monoclonal antibody (McAb). The hapten TTX was linked to carrier protein keyhole limpet hemocyanin (KLH) as immunogen, and linked to ovalbumin (OVA) as coating antigen by the Mannich method. Then the 6~8 weeks Babl/c mice were immunized. After cell amalgamation, a cell line with high specificity and sensitivity was obtained, and McAb was produced. The indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed for detecting TTX. The optimized working conditions of the icELISA were 4.0 μg/mL coating antigen, 0.75 μg/mL McAb, 45 min competitive time, at room temperature (20~25 °C). The IC50value of this method was 24.0 ng/mL, the working ranges were 5.2~107.6 ng/mL, the intra-assay and inter-assay coefficient of variation (CV %) were 4.2 and 4.5, respectively. This investigation will benefit the assay kit development for detecting TTX.

scholarly journals Aspergillus Galactomannan Enzyme-Linked Immunosorbent Assay Cross-Reactivity Caused by Invasive Geotrichum capitatum

2006 ◽  
Vol 44 (9) ◽  
pp. 3432-3434 ◽  
Author(s):  
M. Giacchino ◽  
N. Chiapello ◽  
S. Bezzio ◽  
F. Fagioli ◽  
P. Saracco ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Geotrichum Capitatum

Enzyme-linked immunosorbent assay of autoantibodies reacting with thyroid plasma membrane antigens in sera of patients with autoimmune thyroid diseases

1986 ◽  
Vol 113 (2) ◽  
pp. 255-260 ◽  
Author(s):  
Andrzej Gardas ◽  
Kathleen L. Rives
Keyword(s):  
Plasma Membrane ◽  
Immunosorbent Assay ◽  
Lupus Erythematosus ◽  
Antithyroid Drug ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Autoimmune Thyroid Diseases ◽  
Nodular Goitre ◽  
Membrane Antigens ◽  
Toxic Nodular Goitre

Abstract. A sensitive and specific enzyme-linked immunosorbent assay (ELISA) for the detection of autoantibodies reacting with thyroid plasma membrane antigens has been established. Autoantibodies reacting with thyroid plasma membrane antigens were detected by the ELISA in 95% of untreated hyperthyroid Graves', 68% of antithyroid drug-treated Graves' up to four months of the therapy, in 62% of Hashimoto's thyroiditis and in 8.9% of toxic nodular goitre. The ELISA was negative in 100% healthy blood donors, 100% non-toxic nodular goitre, in 12 patients with rheumatoid arthritis, 18 patients with scleroderma and 94% of patients with systemic lupus erythematosus. The mean value of autoantibodies titre was higher in untreated hyperthyroid Graves' (1:84 000) and lowest in positive patients with autoimmune disease of non-thyroid origin (1:4000). The cross-reactivity of antimicrosomal antigen antibodies was below 10%; there was no influence of antithyroglobulin antibodies on the ELISA; and most of the autoantibodies react with plasma membrane antigens different from the TSH binding sites.

Immunological discrimination between a sap-staining fungus and a biological control fungus

1990 ◽  
Vol 68 (7) ◽  
pp. 1578-1588 ◽  
Author(s):  
Brian T. Luck ◽  
Colette Breuil ◽  
David L. Brown
Keyword(s):  
Electron Microscopy ◽  
Biological Control ◽  
Immunosorbent Assay ◽  
Liquid Culture ◽  
Biological Control Agent ◽  
Dry Weight ◽  
Cross Reactivity ◽  
Immunogold Labeling ◽  
Enzyme Linked Immunosorbent Assay ◽  
Polyclonal Serum

An enzyme-linked immunosorbent assay (ELISA) was used to detect a sap-staining fungus, Ophiostoma piceae, and a biological-control agent, Gliocladium roseum, grown in liquid culture and in wood. A polyclonal serum prepared against whole cell fragments from broken mycelia of O. piceae detected O. piceae in liquid culture at 0.25 μg dry weight/mL; however, there was moderate cross-reactivity with G. roseum. Antiserum adsorbed on G. roseum had almost no reactivity with G. roseum but still reacted strongly with O. piceae. The specificity of these sera was verified, and the antigenic sites were localized, by immunogold labeling and electron microscopy. These studies confirmed that the adsorbed serum could differentiate between G. roseum and O. piceae and showed that the cell wall was the most reactive cellular component. These results are discussed in relation to the development of immunological probes for the detection of sap-staining and biological control fungi. Key words: polyclonal serum, enzyme-linked immunosorbent assay, immunogold labeling, sap-staining and biological control fungi, electron microscopy.

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