A Simple and Rapid Light-Initiated Chemiluminescence Assay for Quantitation of Artemisia-Specific Immunoglobulin E

2021 ◽  
pp. 1-8
Author(s):  
Dandan Liu ◽  
Bei Zhang ◽  
Lina Zhu ◽  
Lisheng Zheng ◽  
Shaoshen Li ◽  
...  
Keyword(s):  
Food Allergen ◽  
Clinical Guideline ◽  
Immunoglobulin E ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Chemiluminescence Assay ◽  
Limit Of Quantitation ◽  
Specific Immunoglobulin E ◽  
Specific Immunoglobulin

<b><i>Background:</i></b> Light-initiated chemiluminescence assay (LICA) is a homogeneous assay that has been successfully used for the quantitation of food allergen-specific immunoglobulin E (sIgE), but not inhaled allergen-sIgE. Simultaneously, current assays used to detect allergen-sIgE are serum consuming and/or time consuming. Hence, we established a method for the quantitation of <i>Artemisia</i>-sIgE based on LICA and verified its performance according to the clinical guideline documents, laying a foundation for the quantitation of inhaled and food allergen-sIgE in parallel on LICA. <b><i>Methods:</i></b> The assay was established after optimizing the first incubation time and the dilutions of <i>Artemisia</i>-coated chemibeads, biotinylated goat anti-human IgE, and serum. In order to quantitate <i>Artemisia</i>-sIgE, the calibration curve was established with a high positive serum of known concentration. The assay performance was confirmed per the clinical guideline documents. In addition, the correlation between the results of LICA and capture enzyme-linked immunosorbent assay was evaluated. <b><i>Results:</i></b> The developed LICA’s coefficients of variation of repeatability and intermediate precision were 3.20%, 2.14%, and 3.85% and 4.30%, 4.00%, and 4.40%, respectively. The limit of detection was 0.10 kU<sub>A</sub>/L, and the limit of quantitation was 0.11 kU<sub>A</sub>/L. The range of linearity was from 0.27 kU<sub>A</sub>/L to 97.53 kU<sub>A</sub>/L (<i>r</i> = 0.9968). The correlation coefficient (<i>r</i>) for the correlation analysis between results of LICA and capture ELISA was 0.9087. This assay was successfully applied in 64 human serum samples, showing good sensitivity (82.20%) and specificity (100%). <b><i>Conclusion:</i></b> An <i>Artemisia</i>-sIgE quantitation assay based on LICA was successfully established. Its performance satisfied the clinical requirements and could be widely used in clinical laboratories.

scholarly journals Prevalence of Allergic Sensitization to Conifer Pollen in a High Cypress Exposure Area

2016 ◽  
Vol 7 (4) ◽  
pp. ar.2016.7.0183 ◽  
Author(s):  
Javier Domínguez-Ortega ◽  
María Ángeles López-Matas ◽  
María Dolores Alonso ◽  
Angélica Feliu ◽  
Javier Ruiz-Hornillos ◽  
...  
Keyword(s):  
Immunoglobulin E ◽  
Skin Prick Testing ◽  
Allergic Sensitization ◽  
Positive Skin Prick Test ◽  
Serum Samples ◽  
Positive Skin ◽  
Prick Test ◽  
Specific Immunoglobulin E ◽  
Specific Immunoglobulin ◽  
Positive Results

Background Sensitization to Finales (Cupressaceae and Pinaceae) has increased dramatically in recent years. The prevalence of sensitization in different geographic areas is related to exposure to specific pollens. Objectives To investigate the prevalence of allergy to different conifer pollens, describe the characteristics of patients with such allergy, and identify the involved allergens. Methods Patients were recruited at five hospitals near Madrid. Extracts from conifer pollen were prepared and used in skin-prick testing. Wheal sizes were recorded, and serum samples obtained from patients with positive reactions to Cupressus arizonica and/or Pinus pinea. The specific immunoglobulin E value to C. arizonica and Cup a 1 was determined. Individual immunoblots for each patient and with a pool of sera were performed. Allergenic proteins were sequenced by using liquid chromatography-tandem mass spectrometry. Results Of 499 individuals included in the study, 17 (14%) had positive skin-prick test results to some conifer pollen extracts. Sixty-four patients had positive results to C. arizonica (prevalence 12.8%) and 11 had positive results to P. pinea (2.2%). All the patients had respiratory symptoms (61.4% during the C. arizonica pollination period), and 62.9% had asthma. Approximately 86% of the patients had positive specific immunoglobulin E results to C. arizonica and 923% had positive results to Cup a 1. Fourteen different bands were recognized by immunoblot; the most frequent bands were those detected at 43, 18, 16, and 14 kDa. All sequenced proteins corresponded to Cup a 1. Conclusion Allergy to conifer pollen could be considered a relevant cause of respiratory allergy in central Spain. Asthma was more frequent than in other studies. We only identified Cup a 1 as involved in sensitization.

scholarly journals Characterization and identification of charcoal of inedible Kerandang fish (Channa pleurophthalmus Blkr) body parts and potential antiallergenic properties

2020 ◽  
Vol 13 (7) ◽  
pp. 1480-1486
Author(s):  
Aryani Aryani ◽  
Eddy Suprayitno ◽  
Bambang Budi Sasmito ◽  
Hardoko Hardoko
Keyword(s):  
Ethyl Acetate ◽  
Immunoglobulin E ◽  
High Resolution Mass Spectrometry ◽  
Ethyl Acetate Extract ◽  
The Body ◽  
Enzyme Linked Immunosorbent Assay ◽  
Body Parts ◽  
Specific Immunoglobulin E ◽  
Specific Immunoglobulin ◽  
Resolution Mass

Background and Aim: The study about the antiallergenic properties of inedible fish body parts is still limited. Therefore, this study aimed to characterize the charcoal from the body parts of Kerandang fish (Channa pleurophthalmus Blkr) and identify its antiallergenic properties. Materials and Methods: This study used some non-edible body parts extracted from the Kerandang fish (i.e., the scalp, scales, and dorsal, pectoral, ventral, anal, and caudal fins) using a maceration method with different solvents (ethanol, ethyl acetate, and chloroform). The identification of active compounds in the extract was carried out using liquid chromatography– high-resolution mass spectrometry (LC-HRMS) analysis, while the antihyaluronidase activity was determined using the antihyaluronidase test. The highest charcoal antihyaluronidase activity-extract was applied to ovalbumin-induced mice for 7 days with various doses (10, 15, and 20 mg/kg). The specific immunoglobulin E (IgE) was measured using enzyme-linked immunosorbent assay on day 8. Results: Our LC-HRMS analysis showed that the active compound of charcoal in the caudal fins of Kerandang fish was hexadecanamide. The highest inhibition (IC50) of hyaluronidase was found in the ethyl acetate extract of fish caudal fins at a concentration of 4 mg/mL. We found that 15 mg/kg body weight of charcoal of fish caudal fins suppressed IgE expression in male mice. Conclusion: Our findings indicate that the charcoal of non-edible body parts of Kerandang and one of its constituent, hexadecanamide, may have strong antiallergic effects.

scholarly journals Proficiency monitoring of monoclonal antibody cocktail–based enzyme-linked immunosorbent assay for detection of allergen-specific immunoglobulin E in dogs

2015 ◽  
Vol 27 (4) ◽  
pp. 461-469 ◽  
Author(s):  
Kenneth W. Lee ◽  
Karen Blankenship ◽  
Brennan McKinney ◽  
Gerhard Kern ◽  
Jesse Buch ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Immunoglobulin E ◽  
Enzyme Linked Immunosorbent Assay ◽  
Negative Results ◽  
Specific Immunoglobulin E ◽  
Specific Immunoglobulin ◽  
Density Values ◽  
Response Profiles ◽  
Antibody Cocktail

The purpose of our study was to document the continued comparative proficiency of different laboratories that perform a monoclonal antibody–based enzyme-linked immunosorbent assay (macELISA) for detection of allergen-specific immunoglobulin (Ig)E in dogs. Replicate samples of 18 different sera pools were independently evaluated in a single blinded fashion by each of 16 different operators functioning in 10 different laboratories. The average intra-assay variance among reactive assay calibrators in all laboratories was 6.0% (range: 2.7–16.1%), while the average intralaboratory interassay variance was 7.5% (range: 3.9–10.9%). The overall interassay interlaboratory variance was consistent among laboratories and averaged 11.4% (range: 8.5–12.5%). All laboratories yielded similar profiles and magnitudes of responses for replicate unknown samples; dose response profiles observed in each of the laboratories were indistinguishable. Considering the positive or negative results, interassay interlaboratory concordance of results exceeded 90%. Correlation of optical density values between and among all laboratories was strong ( r > 0.9, P < 0.001). Collectively, the results demonstrated that the macELISA for measuring allergen-specific canine IgE is reproducible, and documents that consistency of results can be achieved not only in an individual laboratory by differing operators but also among laboratories using the same monoclonal-based ELISA.

scholarly journals Cross-reactivity of each fraction among cereals in children with wheat allergy

2021 ◽  
Author(s):  
Mari Takei ◽  
Akemi Saito ◽  
Noriyuki Yanagida ◽  
Sakura Sato ◽  
Motohiro Ebisawa
Keyword(s):  
Immunoglobulin E ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Wheat Allergy ◽  
Protein Levels ◽  
Inhibition Elisa ◽  
Specific Immunoglobulin E ◽  
Specific Immunoglobulin ◽  
Lower Protein

Background: Cross-reactivity between wheat and other cereals is an essential issue in the management of wheat allergy. Few studies have reported in vitro cross-reactivity in immediate-type wheat allergy. This study aimed to examine cross-reactivity of the three fractions (albumin/globulin, gliadin, and glutenin fractions) among cereals in children with wheat allergy. Methods: Sera from 128 children with immediate-type wheat allergy were collected. We measured specific immunoglobulin E (sIgE) levels against each fraction of wheat, barley, and rye by using an enzyme-linked immunosorbent assay (ELISA). Cross-reactivities of each fraction among wheat, barley, and rye were examined via inhibition ELISA. Results: All subjects were sensitized to all the fractions of wheat, and also those of barley and rye. The wheat sIgE levels were significantly higher than those of barley and rye in all the fractions (p ≤ 0.001) and were significantly correlated with sIgE levels to them in each fraction (r = 0.887–0.969, p < 0.001). On inhibition ELISA, wheat inhibited the IgE binding to most of the solid phases at the lower protein levels compared to barley and rye in all fractions. Conclusions: In children with immediate-type wheat allergy, sensitization to all the three fractions of wheat was observed. In addition, they showed sensitization to barley and rye caused by in vitro cross-reactivity with wheat in each fraction. When managing children with wheat allergy, sensitization to barley and rye caused by the cross-reactivities should be considered.

scholarly journals Serum Immunoglobulin E Levels in Patients with Allergic Conjunctivitis and Contact Lens Wearers

1970 ◽  
Vol 12 (3) ◽  
pp. 152-155
Author(s):  
Ozlem Gurses Sahin ◽  
Nusret Taheri
Keyword(s):  
Contact Lens ◽  
Allergic Conjunctivitis ◽  
Immunoglobulin E ◽  
Total Duration ◽  
Serum Immunoglobulin ◽  
Total Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Immunoglobulin E ◽  
Specific Immunoglobulin

Aim: To determine the differences between total and serum-specific immunoglobulin E levels in patients with type 1 allergic conjunctivitis and asymptomatic contact lens wearers. The correlation of total serum immunoglobulin E level of asymptomatic contact lens wearers with contact lens wearing time, and total duration of contact lens use was also evaluated.Methods: This was a case-control study involving 25 asymptomatic contact lens wearers, 25 patients with type 1 allergic conjunctivitis, and 25 age- and sex-matched healthy controls. Total serum immunoglobulin E levels were detected by enzyme-linked immunosorbent assay. Serum-specific IgE analysis against the listed indoor, food, and outdoor allergens were studied by immunofluorescence assay for participants whose total serum immunoglobulin E levels were >100 IU/mL. Pearson’s and Spearman’s correlations were used for bivariate analysis. Statistical significance was accepted at the 0.05 level.Results: The mean level of total serum immunoglobulin E was greater for patients with type 1 allergic conjunctivitis than for contact lens wearers and controls. Serum-specific immunoglobulin E detected in patients with type 1 allergic conjunctivitis was against indoor, food, and outdoor allergens, while serum-specific immunoglobulin E detected in contact lens wearers was only against outdoor allergens. A statistically significant correlation was found for total serum immunoglobulin E levels of contact lens wearers with contact lens wearing time.Conclusions: These results suggest that differences in serum total and specific immunoglobulin E levels exist between patients with type 1 allergic conjunctivitis and CL wearers and controls. Further research in a larger group of patients is needed to validate these findings.

Sign in / Sign up

Export Citation Format

Share Document