Reproducibility of antigen-immobilized enzyme-linked immunosorbent assay (ELISA) and sandwich ELISA for quantitative detection of NNV particles

2020 ◽  
Vol 275 ◽  
pp. 113754 ◽  
Author(s):  
Hyun Jung Gye ◽  
Toyohiko Nishizawa
Keyword(s):  
Immunosorbent Assay ◽  
Immobilized Enzyme ◽  
Sandwich Elisa ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay

scholarly journals Development of a Smartphone-Based Nanozyme-Linked Immunosorbent Assay for Quantitative Detection of SARS-CoV-2 Nucleocapsid Phosphoprotein in Blood

2021 ◽  
Vol 12 ◽  
Author(s):  
Bochao Liu ◽  
Ze Wu ◽  
Chaolan Liang ◽  
Jinhui Lu ◽  
Jinfeng Li ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Cross Reactivity ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Primary Screening ◽  
Global Pandemic ◽  
Novel Coronavirus ◽  
Acid Test ◽  
Control Samples

Since December 2019, a novel coronavirus (SARS-CoV-2) has resulted in a global pandemic of coronavirus disease (COVID-19). Although viral nucleic acid test (NAT) has been applied predominantly to detect SARS-CoV-2 RNA for confirmation diagnosis of COVID-19, an urgent need for alternative, rapid, and sensitive immunoassays is required for primary screening of virus. In this study, we developed a smartphone-based nanozyme-linked immunosorbent assay (SP-NLISA) for detecting the specific nucleocapsid phosphoprotein (NP) of SARS-CoV-2 in 37 serum samples from 20 COVID-19 patients who were diagnosed by NAT previously. By using SP-NLISA, 28/37 (75.7%) serum samples were detected for NP antigens and no cross-reactivity with blood donors’ control samples collected from different areas of China. In a control assay using the conventional enzyme-linked immunosorbent assay (ELISA), only 7/37 (18.91%) serum samples were detected for NP antigens and no cross-reactivity with control samples. SP-NLISA could be used for rapid detection of SARS-CoV-2 NP antigen in primary screening of SARS-CoV-2 infected individuals.

Detection of circulating antigen in bancroftian filariasis by sandwich ELISA using filarial serum IgG

1984 ◽  
Vol 58 (3) ◽  
pp. 259-262 ◽  
Author(s):  
M. V. R. Reddy ◽  
Ashok Malhotra ◽  
B. C. Harinath
Keyword(s):  
Immunosorbent Assay ◽  
Sandwich Elisa ◽  
Negative Reaction ◽  
Serum Immunoglobulin ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bancroftian Filariasis ◽  
Positive Correlation ◽  
Serum Igg ◽  
Filarial Antigen ◽  
Circulating Antigen

ABSTRACTThe utility of the IgG fraction of human filarial serum immunoglobulin in detecting circulating antigen by sandwich enzyme linked immunosorbent assay (ELISA) was studied. 27 of 33 sera from persons with microfilaracmia, 19 of 30 sera from clinical cases of filariasis, 4 of 30 sera from normal persons from a region endemic for filariasis showed the presence of circulating filarial antigen. All the 20 normal sera from the area where filariasis was not endemic gave negative reaction for filarial antigen. Those sera from persons with microfilaracmia that showed the presence of circulating antigen also showed an apparent positive correlation between the microfilarial density and the antigen titre.

scholarly journals A Portable, Cost-Effective and User-Friendly Instrument for Colorimetric Enzyme-Linked Immunosorbent Assay and Rapid Detection of Aflatoxin B1

Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2483
Author(s):  
Wenzhi Tang ◽  
Yangchun Qi ◽  
Zhonghong Li
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Rapid Detection ◽  
Attractive Potential ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ambient Light ◽  
User Friendliness ◽  
Signal Light ◽  
User Friendly

Food analysis based on the enzyme-linked immunosorbent assay (ELISA) is simple, sensitive and rapid, but requires a costly colorimetric instrument. The aim of this work was to develop a portable, low-cost and user-friendly colorimetric instrument for colorimetric ELISA and aflatoxin B1 (AFB1) detection. The principle of the developed instrument was employing a light-emitting diode to generate the signal light and using a light-dependent resistor to measure the signal light absorbed by the oxidized 3,3′,5,5′-tetramethyl benzidine. The absorption spectra revealed that the solution absorbed signal light more strongly after reaction with H2SO4, and blue light would be favorably absorbed. Evaluations on the stability and accuracy of the instrument and interference from ambient light showed that the fabricated instrument was stable, accurate, capable of quantitative detection and insensitive to ambient light changes. In addition, this instrument is user-friendly since it could calculate and report the final amount of AFB1 to the operator. Measurements of maize and peanuts showed that the instrument provided as accurate results as the professional equipment. With the low fabrication cost (about RMB 129 or USD 20), portability, and user-friendliness, this instrument presents attractive potential in the rapid detection of AFB1.

Sandwich ELISA for Detection of Pig Meat in Raw Beef Using Antisera to Muscle Soluble Proteins

1988 ◽  
Vol 51 (10) ◽  
pp. 790-798 ◽  
Author(s):  
ROSARIO MARTÍN ◽  
JUAN I. AZCONA ◽  
CARMEN CASAS ◽  
PABLO E. HERNÁNDEZ ◽  
BERNABÉ SANZ
Keyword(s):  
Horseradish Peroxidase ◽  
Immunosorbent Assay ◽  
Peroxidase Activity ◽  
Sandwich Elisa ◽  
Soluble Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Double Antibody ◽  
Colored Product ◽  
Pig Meat

A double-antibody sandwich ELISA (enzyme-linked immunosorbent assay) has been successfully developed for the detection of defined amounts of pig meat (1–50%) in raw beef. Antibodies against pig sarcoplasmic extracts were produced in rabbits. Pig-specific antibodies were affinity purified by removing antibodies which crossreacted with horse, chicken or beef extracts followed by immunoadsorption and elution from a pig-extract column. The ELISA involved capturing antigens in sarcoplasmic extracts with pig specific antibodies immobilized on 96-well plates, detecting bound antigen with pig specific, horseradish peroxidase-labeling antibody, and measuring peroxidase activity by the conversion of a clear substrate to a colored product.

scholarly journals Evaluation of a Commercial Sandwich Enzyme-Linked Immunosorbent Assay for the Quantification of Beta-Casomorphin 7 in Yogurt Using Solid-Phase Extraction Coupled to Liquid Chromatography-Tandem Mass Spectrometry as the “Gold Standard” Method

2018 ◽  
Vol 101 (2) ◽  
pp. 515-519 ◽  
Author(s):  
Duc Doan Nguyen ◽  
Francesco Busetti ◽  
Stuart Keith Johnson ◽  
Vicky Ann Solah
Keyword(s):  
Immunosorbent Assay ◽  
Standard Method ◽  
Gold Standard ◽  
Solid Phase ◽  
Sandwich Elisa ◽  
Aminopeptidase Activity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gold Standard Method ◽  
Liquid Chromatography Tandem Mass ◽  
Dipeptidyl Aminopeptidase

Abstract This study investigated beta-casomorphin 7 (BCM7) in yogurt by means of LC-tandem MS (MS/MS) and enzyme-linked immunosorbent assay (ELISA) and use LC-MS/MS as the “gold standard” method to evaluate the applicability of a commercial ELISA. The level of BCM7 in milk obtained from ELISA analysis was much lower than that obtained by LC-MS/MS analysis and trended to increase during fermentation and storage of yogurt. Meanwhile, the results obtained from LC-MS/MS showed that BCM7 degraded during stages of yogurt processing, and its degradation may have been caused by X-prolyl dipeptidyl aminopeptidase activity. As a result, the commercial sandwich ELISA kit was not suitable for the quantification of BCM7 in fermented dairy milk.

scholarly journals Detection of Plasmodium Aldolase Using a Smartphone and Microfluidic Enzyme Linked Immunosorbent Assay

2017 ◽  
Vol 2017 ◽  
pp. 1-4
Author(s):  
Nikhil S. Gopal ◽  
Ruben Raychaudhuri
Keyword(s):  
Immunosorbent Assay ◽  
Microfluidic Chip ◽  
Rural Areas ◽  
Color Change ◽  
Low Cost ◽  
Sandwich Elisa ◽  
Microfluidic System ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cost System ◽  
Therapeutic Outcomes

Background. Malaria control efforts are limited in rural areas. A low-cost system to monitor response without the use of electricity is needed. Plasmodium aldolase is a malaria biomarker measured using enzyme linked immunosorbent assay (ELISA) techniques. A three-part system using ELISA was developed consisting of a microfluidic chip, hand crank centrifuge, and a smartphone. Methods. A circular microfluidic chip was fabricated using clear acrylic and a CO2 laser. A series of passive valves released reagents at precise times based upon centrifugal force. Color change was measured via smartphone camera using an application programmed in Java. The microchip was compared to a standard 96-well sandwich ELISA. Results. Results from standard ELISA were compared to microchip at varying concentrations (1–10 ng/mL). Over 15 different microfluidic patterns were tested, and a final prototype of the chip was created. The prototype microchip was compared to standard sandwich ELISA (n=20) using samples of recombinant aldolase. Color readings of standard ELISA and microfluidic microchip showed similar results. Conclusion. A low-cost microfluidic system could detect and follow therapeutic outcomes in rural areas and identify resistant strains.

A Competitive Enzyme-Linked Immunosorbent Assay for Detection of Bovine Milk in Ovine and Caprine Milk and Cheese Using a Monoclonal Antibody against Bovine β-Casein

1997 ◽  
Vol 60 (1) ◽  
pp. 64-66 ◽  
Author(s):  
G. ANGUITA ◽  
R. MARTÍN ◽  
T. GARCÍA ◽  
P. MORALES ◽  
A. I. HAZA ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Binding Sites ◽  
Bovine Milk ◽  
Microtiter Plate ◽  
Competitive Elisa ◽  
Quantitative Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Caprine Milk ◽  
Mouse Immunoglobulin

A competitive ELISA (enzyme-linked immunosorbent assay) was performed to detect and quantify bovine milk in ovine and caprine milk and cheese using a monoclonal antibody (AH4 MAb) against bovine β-casein. Ovine or caprine milk and cheese containing bovine milk were added simultaneously with the AH4 MAb to the wells of a microtiter plate that had been previously sensitized with commercial bovine β-casein. The bovine caseins in milk or cheese samples compete with the bovine β-casein bound to the plate for the AH4 MAb binding sites. Further immunorecognition of AH4 MAb bound to the bovine β-casein immobilized onto the plate was attained with rabbit anti-mouse immunoglobulin conjugated to peroxidase. Subsequent enzymic conversion of the substrate showed clear differences in absorbance values during assay of mixtures of ovine and caprine milk and cheese containing various amounts of bovine milk. The competitive ELISA developed in this work allows the quantitative detection of bovine milk in ovine and caprine milk and cheese samples in the range of 0.5 to 25% of substitution.

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