scholarly journals Determination of Aspergillus Fumigatus Allergen 1 in Poultry Farms Using the Enzyme Immunoassay

2010 ◽  
Vol 61 (2) ◽  
pp. 167-173 ◽  
Author(s):  
Ljerka Prester ◽  
Jelena Macan ◽  
Kristina Matković ◽  
Marija Vučemilo
Keyword(s):  
Aspergillus Fumigatus ◽  
Enzyme Immunoassay ◽  
Limit Of Detection ◽  
Major Allergen ◽  
Indoor Dust ◽  
Enzyme Linked Immunosorbent Assay ◽  
Poultry Farms ◽  
Asp F 1 ◽  
Common Genus

Determination ofAspergillus FumigatusAllergen 1 in Poultry Farms Using the Enzyme ImmunoassayPoultry farms contain high levels of allergenic fungi, andAspergillusspp. is the most common genus of moulds.Aspergillus fumigatusantigens are responsible for the development of several respiratory diseases including asthma. The aim of this study was to measure the mass fraction of Asp f 1, a major allergen ofAsperillus fumigatusin 37 indoor dust samples collected from four poultry farms in a rural area of the Zagreb County (Croatia) using the enzyme-linked immunosorbent assay. More than 62 % of dust samples had detectable Asp f 1 levels (limit of detection 3.6 ng g-1). The overall mean Asp f 1 level was 17.9 ng g-1[range (3.8 to 72.4) ng g-1]. Satisfactory results were obtained for analytical within-run imprecision (6.7 %), between-run imprecision (10.5 %), and accuracy (91 % to 115 %). Microclimate parameters (air temperature, relative humidity, and velocity) were within the recommended ranges in all poultry farms. This study has shown that Asp f 1 settles on dust at poultry farms and that occupational exposure to this allergen deserves monitoring in livestock buildings.

scholarly journals Pitfalls in the Immunochemical Determination of β‑Lactam Antibiotics in Water

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 298
Author(s):  
Alexander Ecke ◽  
Rudolf J. Schneider
Keyword(s):  
Limit Of Detection ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Degree Of Hydrolysis ◽  
Site Analysis ◽  
Antibody Recognition ◽  
Immunochemical Determination ◽  
Key Factor ◽  
Lactam Ring

Contamination of waters with pharmaceuticals is an alarming problem as it may support the evolution of antimicrobial resistance. Therefore, fast and cost-effective analytical methods for potential on-site analysis are desired in order to control the water quality and assure the safety of its use as a source of drinking water. Antibody-based methods, such as the enzyme-linked immunosorbent assay (ELISA), can be helpful in this regard but can also have certain pitfalls in store, depending on the analyte. As shown here for the class of β-lactam antibiotics, hydrolysis of the β‑lactam ring is a key factor in the immunochemical analysis as it influences antibody recognition. With the antibody used in this study, the limit of detection (LOD) in the immunoassay could be significantly reduced by hydrolysis for the five tested penicillins, with the lowest LOD for carbenicillin (0.2 nmol/L) and the greatest impact on penicillins G and V (reduction by 85%). In addition to enhanced quantification, our strategy also provides access to information about the degree of hydrolysis in water samples as shown for the most abundant penicillin amoxicillin.

scholarly journals Characterization and validation of fragment antigen-binding (Fab) antibody-based immunoassay for deoxymiroestrol, a potent phytoestrogen from Pueraria candollei

2021 ◽  
Author(s):  
Worapol Sae-foo ◽  
Supaluk Krittanai ◽  
Wipawee Juengsanguanpornsuk ◽  
Gorawit Yusakul ◽  
Tharita Kitisripanya ◽  
...  
Keyword(s):  
Quantitative Method ◽  
Estrogenic Activity ◽  
Limit Of Detection ◽  
Hormone Replacement ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antigen Binding ◽  
Pueraria Candollei ◽  
Specific Determination ◽  
Fragment Antigen Binding

Deoxymiroestrol is the most potent phytoestrogen in chromenes group that has been found in Pueraria candollei, Thai name known as Kwao Krua Khao. Several studies reported estrogenic activity of P. candollei in order to using as hormone replacement therapy for postmenopausal women. Previously, specific determination of deoxymiroestrol content by enzyme-linked immunosorbent assay (ELISA) using polyclonal antibody (pAb) have been reported. However, production of pAb has limitation and variability from different batches. Therefore, in this study, we established quantitative method for determination of deoxymiroestrol using fragment antigen-binding (Fab) antibody-based immunoassay. The developed immunoassay has specificity to deoxymiroestrol with a calibration range of 15.6-1000 ng mL-1. Precision including intra-assay and inter-assay are 1.48-7.11 and 0.58-9.31%, respectively. Accuracy of the assay showed in recovery between 99.77-101.61% when spike deoxymiroestrol standard into the samples. The limit of detection (LOD) is 30.80 ng mL-1. Comparation antibody-based immunoassay for determination of deoxymiroestrol using Fab with pAb was represented consistency (R2 = 0.9807) when analysis roots bark of Pueraria candollei from difference areas. Therefore, this development assay can apply to determine deoxymiroestrol content in the plant samples.

A Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Determination of Homoharringtonine

Planta Medica ◽  
2018 ◽  
Vol 84 (14) ◽  
pp. 1038-1044 ◽  
Author(s):  
Benyakan Pongkitwitoon ◽  
Seiichi Sakamoto ◽  
Rika Nagamitsu ◽  
Waraporn Putalun ◽  
Hiroyuki Tanaka ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Myeloid Leukemia ◽  
High Performance ◽  
Sodium Periodate ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Study Support ◽  
Mediated Oxidation

AbstractHomoharringtonine (HHT), also known as omacetaxine, is a natural compound found in the genus Cephalotaxus and is a promising pharmaceutical drug used for the treatment of chronic or accelerated phase chronic myeloid leukemia. As a tool for the quantitative determination of HHT, a specific monoclonal antibody against HHT (MAb 6A1) was generated by conjugates prepared via sodium periodate-mediated oxidation. The developed indirect competitive enzyme-linked immunosorbent assay (icELISA) using MAb 6A1 was found to be highly specific and sensitive with a limit of detection for HHT of 48.8 ng/mL. Validation assays to evaluate precision and accuracy of the method were conducted by the use of intra- and inter-assay analysis, recovery test, and comparison analysis between the amounts of HHT determined by ELISA and high-performance liquid chromatography. These results revealed that the established icELISA using MAb 6A1 is specific, sensitive, and reliable enough to be applied to the qualitative analysis for HHT. Furthermore, the results of this study support the usefulness of sodium periodate as a reagent for the conjugation between Cephalotaxus alkaloids and proteins for producing specific antibodies.

scholarly journals Use of a Synthetic Peptide Epitope of Asp f 1, a Major Allergen or Antigen of Aspergillus fumigatus, for Improved Immunodiagnosis of Allergic Bronchopulmonary Aspergillosis

2004 ◽  
Vol 11 (3) ◽  
pp. 552-558 ◽  
Author(s):  
Taruna Madan ◽  
Priyanka Priyadarsiny ◽  
Mudit Vaid ◽  
Neel Kamal ◽  
Ashok Shah ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Synthetic Peptide ◽  
Immunoglobulin E ◽  
Allergic Bronchopulmonary Aspergillosis ◽  
Major Allergen ◽  
Specific Ige ◽  
Diagnostic Efficiency ◽  
Intradermal Testing ◽  
Peptide Epitope ◽  
Asp F 1

ABSTRACT Allergic bronchopulmonary aspergillosis (ABPA) is an immunologically complex allergic disorder caused by the fungal pathogen Aspergillus fumigatus. Elevated levels of total immunoglobulin E (IgE), specific IgE, and IgG antibodies in sera are important immunodiagnostic criteria for ABPA. International reference standards or standardized immunodiagnostic assays are not available due to a lack of well-defined diagnostic antigens. The present study was carried out to identify and evaluate the immunodiagnostic relevance of synthetic epitopic peptides of Asp f 1, a major allergen, antigen, or cytotoxin of A. fumigatus. Five overlapping peptides were synthesized from the N terminus of Asp f 1, one of the potential immunodominant regions predicted by algorithmic programs. The 11-amino-acid synthetic peptide (P1) significantly inhibited both IgG binding (89.10% ± 4.45%) and IgE binding (77.32% ± 3.38%) of the standardized diagnostic antigen (SDA) (a well-defined pool of diagnostically relevant allergens and antigens of A. fumigatus). With a panel of sera of ABPA patients, allergic patients with skin test negativity to A. fumigatus, and healthy individuals, P1 showed a higher diagnostic efficiency than SDA (specific IgG, 100%; specific IgE, 98.3%). The diagnostic efficiency of P1 could be attributed to the presence of homologous epitopes in various immunodominant allergens or antigens of A. fumigatus. The ability of P1 to induce histamine release from sensitized mast cells and a Th2 type of cytokine profile in peripheral blood mononuclear cells of ABPA patients suggests its potential for use in intradermal testing. P1 could be further explored for development of a standardized, specific, and sensitive immunodiagnostic test for aspergillosis.

Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody

Planta Medica ◽  
2017 ◽  
Vol 83 (10) ◽  
pp. 855-861 ◽  
Author(s):  
Tharita Kitisripanya ◽  
Supaluk Krittanai ◽  
Orapin Udomsin ◽  
Kamonthip Jutathis ◽  
Jukrapun Komaikul ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Estrogenic Activity ◽  
Limit Of Detection ◽  
Simple Procedure ◽  
Enzyme Linked Immunosorbent Assay ◽  
Efficacy And Safety ◽  
Limit Of Quantitation ◽  
Estrogenic Action ◽  
White Kwao Krua

AbstractMiroestrol is a chromene with potent estrogenic activity present in Pueraria candollei, commonly known as White Kwao Krua. Although this compound is only present in low amounts in the plant, it plays an important role in the estrogenic action of P. candollei products. As a tool for further studies about the efficacy and safety of P. candollei as a phytoestrogenic supplement, we generated a novel monoclonal antibody against miroestrol. This anti-miroestrol monoclonal antibody was used to develop an immunoassay for the determination of miroestrol content, which can be used for quality control purposes of P. candollei. The developed ELISA against miroestrol has a calibration range of 10–780 ng/mL miroestrol, a limit of detection of 3.5 ng/mL, and a limit of quantitation of 12.2 ng/mL. According to the validation analysis, the established ELISA is precise, accurate, specific, and sensitive for miroestrol detection in plants. Furthermore, the anti-miroestrol monoclonal antibody was used to prepare an immunoaffinity column for the isolation of miroestrol from the tuberous root of P. candollei. The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel.

scholarly journals Quantitative analysis of chloramphenicol residues in shrimp muscle tissues by Chemiluminescent enzyme immunoassay

2011 ◽  
Vol 23 (No. 6) ◽  
pp. 251-256
Author(s):  
X. Chuanlai ◽  
P. Cifang ◽  
H. Kai ◽  
J. Zhengyu ◽  
W. Wukang
Keyword(s):  
Enzyme Immunoassay ◽  
Limit Of Detection ◽  
Tween 20 ◽  
Detection Range ◽  
Muscle Tissues ◽  
Physico Chemical ◽  
Interassay Coefficient ◽  
Chemiluminescent Enzyme Immunoassay ◽  
Better Than

A competitive indirect chemiluminescent enzyme immunoassay (ic-CLEIA) has been developed for the determination of chloramphenicol (CAP) residues in shrimp. After the optimisation of four physico-chemical parameters, i.e. incubation time, concentration of Tween-20, concentration of PBS and its pH, the method developed gave a limit of detection of 0.01 ng/ml and a detection range from 0.03 ng/ml to 23.7 ng/ml, with an ED<sub>50</sub> of 0.47 ng/ml. The developed method has been validated on spiked shrimp samples in terms of precision (intra- and interassay coefficient variations of less than 10% and 15%, respectively), and of accuracy (mean recovery from 95% to 123%). All these parameters being better than those of the ELISA method which is widely used to detect chloramphenicol, it may be suggested that the CLEIA method can be used to detect aquatic samples instead of ELISA. &nbsp;

scholarly journals Single-Laboratory Validation of the Biosense Direct Competitive Enzyme-Linked Immunosorbent Assay (ELISA) for Determination of Domoic Acid Toxins in Shellfish

2007 ◽  
Vol 90 (4) ◽  
pp. 1000-1010 ◽  
Author(s):  
Hans Kleivdal ◽  
Sven-Inge Kristiansen ◽  
Mona V Nilsen ◽  
Lyn Briggs
Keyword(s):  
Immunosorbent Assay ◽  
Domoic Acid ◽  
Matrix Effects ◽  
Limit Of Detection ◽  
Method Comparison ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Shellfish Poisoning ◽  
Relative Standard

Abstract Method validation was conducted for an enzyme-linked immunosorbent assay (ELISA) for the determination of domoic acid (DA) toxins, known to give amnesic shellfish poisoning (ASP) symptoms, in shellfish. The calibration curve range of the assay is approximately 10260 pg/mL, with a dynamic working range for DA toxins in shellfish from 0.01 to at least 250 mg/kg. The ASP ELISA showed no significant cross-reactivity to structural analogs, and proved to be robust to deliberate alterations of the optimal running conditions. The shellfish matrix effects observed with mussels, oysters, and scallops were eliminated by diluting shellfish extracts 1:200 prior to analysis, leading to a limit of detection at 0.003 mg/kg. Thirteen blank shellfish homogenates were spiked with certified mussel material containing DA to levels in the range of 0.125 mg DA/kg, and analyzed in quadruplicate on 3 different days. The relative standard deviation (RSD) under intra-assay repeatability conditions ranged from 6.5 to 13.1%, and under interassay repeatability conditions the RSD ranged from 5.7 to 13.4%, with a mean value of 9.3%. The recoveries ranged from 85.5 to 106.6%, with a mean recovery of 102.2%. A method comparison was conducted with liquid chromatography with ultraviolet detection, using naturally contaminated scallop samples (n = 27) with DA levels at 0244 mg/kg. The overall correlation coefficient was 0.960 and the slope of the regression was 1.218, indicating a good agreement between the methods.

Detection of bisphenol A in water samples using ELISA determination method

2011 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
J. Zheng ◽  
S. Q. Zhao ◽  
X. T. Xu ◽  
K. Zhang
Keyword(s):  
Drinking Water ◽  
Bisphenol A ◽  
Water Samples ◽  
Limit Of Detection ◽  
Industrial Effluent ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Uv Spectrophotometry ◽  
Inhibition Concentration

In order to study whether bisphenol A (BPA) can pass into drinking water from polycarbonate barrel and exist in the river and industrial effluent the indirect competitive enzyme-linked immunosorbent assay (ELISA) for the determination of BPA was established. The results presented an inhibition concentration at 50% absorbance (IC50) of 0.123 mg L−1, and the limit of detection (LOD) is 9.934 μg L−1. The specificity of antiserum was proved well because the cross-reactivity with benzene, tert-butylbenzene, hydroquinone and o-hydroxybenzoic acid were found lower than 0.01%, except phenol was 0.26%. The method was found to be reliable and repeatable. It was used for monitoring the concentration of BPA in the barreled drinking water. The results confirmed BPA can pass into barreled drinking water from the polycarbonate barrel and concentration increased as days went on. A certain content of BPA was found in industrial effluent. The results of ELISA were consistent with the results of UV spectrophotometry. BPA could not be found in the water samples obtained from Zhujiang River. The established method shows specific recognition of BPA and could be applied in detection of environmental BPA.

scholarly journals Determination of Egg Proteins in Food Products by Enzyme Immunoassay

2000 ◽  
Vol 83 (1) ◽  
pp. 139-143 ◽  
Author(s):  
Jupiter M Yeung ◽  
W Harvey Newsome ◽  
Michael A Abbott
Keyword(s):  
Detection Limit ◽  
Enzyme Immunoassay ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antigen Binding ◽  
Food Ingredients ◽  
Coefficients Of Variation ◽  
Egg Products ◽  
Whole Egg

Abstract An enzyme-linked immunosorbent assay (ELISA) was developed to determine the presence of egg proteins in foods. The polyclonal antibodies developed were specific to whole egg proteins and did not cross-react with any of the 38 nuts, legumes, or other common food ingredients tested. The concentrations of egg proteins that will inhibit 50% of antibody–antigen binding, IC50, were 3–7 ng/mL, and the linear range was 0.5–62.5 ng/mL. The detection limit was 0.2 ppm for various foods. Recoveries ranged from 67 to 96%. The intra- and inter-assay coefficients of variation in this procedure were 10–13% for ice cream spiked at 0.8 and 1.6 ppm. The ELISA has been applied to ice creams, noodles, pasta, and breads. Egg proteins were identified in all declared egg products, and no false positives were found.

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