Detection Of Immunoglobulin Research Articles

Optical Label-Free Aptasensor Based on Weak Value Amplification for Real-Time and Ultrasensitive Detection of IgE.

Allergy is a prevalent disease, and the potential allergic population is expanding with industrialization and changes in people's living standards. Serum immunoglobulin E (IgE) level is one of the critical indicators for determining allergy. Here, we proposed a simple, real-time monitoring, low chip cost, label-free aptamer biosensing strategy based on weak value amplification (WVA) for the quantitative detection of IgE in serum samples, enabling early and accurate diagnosis of allergic or hypersensitive patients. The aptasensor combined an imaging weak measurement system with the high specificity of the aptamer for the marker IgE. By modifying the amino group at the 3-terminal end, the anti-IgE aptamers can attach to a dopamine-modified prism's surface and selectively recognize IgE in human serum. In the presence of IgE, a specific binding reaction occurred, resulting in a change in the refractive index of the reactive region's surface, manifested as a change in the light intensity of the camera acquired experimental images. As the concentration of IgE increased, the relative light intensity advanced sequentially. The WVA-aptasensing strategy achieved a wide detection range of 0.01 ng/mL to 2 μg/mL in phosphate buffered saline buffer, with the resolution as low as 4.3 pg/mL. IgE testing experiments in human serum have proved the feasibility of our methods in detecting complex samples. In addition, the method specifically recognized IgE without interference from other proteins. We believe that our proposed sensing strategy opens up new possibilities for ultrahigh sensitivity screening of IgE and can be expanded to detecting other biomolecules.

Analysis of Specific IgE Distribution Characteristics of Dust Mite Allergen Components in Patients With Atopic Dermatitis

Objective: To investigate the distribution characteristics of specific immunoglobulin E (IgE) to dust mite components in patients with atopic dermatitis (AD), aiming to provide a reference basis for the development of microarray protein chips for allergen component discrimination diagnosis and personalized desensitization therapy.Methods: A total of 55 patients who visited Huishan District People’s Hospital of Wuxi City, Xishan People’s Hospital of Wuxi City, and People’s Hospital of Wuxi City from January 2023 to December 2023 were selected. The results of serum total IgE (total IgE) and specific IgE detection were analyzed.Results: Among the 55 patients, 54 (98.2%) were positive (cutoff value > 0.35 Ua/mL, the normal value for serum total IgE is under 150–400 IU/mL) for serum total IgE antibodies, 52 (94.6%) were positive for Dermatophagoides pteronyssinus (Dp)–specific IgE antibodies, and 50 (90.9%) were positive for Dermatophagoides farinae (Df)–specific IgE antibodies. The protein chip results were consistent with the existing Phadia detection results.Conclusion: The development of chips based on CRD (component‐resolved diagnosis) cannot only rely on IgE binding rate to assist in precise treatment of patients but also choose personalized desensitization therapy, providing more practical recommendations for preventing cross‐allergy, avoiding allergic environments, or ingesting sensitizing foods for susceptible individuals.

A fluorescence point-of-care testing system for sample-to-answer detection of immunoglobulin E

In the work, a novel fluorescence point-of-care testing (POCT) system was developed for sample-to-answer detection of immunoglobulin E (IgE), which is the first time that a cloth material has been applied to the fluorescence detection of proteins. The system included an lateral/vertical flow-based microfluidic (LVFM) model, POCT detector, and smartphone-based data processing module. In the LVFM model, the non-woven cloth and screen-printing methods were applied to break the restrictions of traditional lateral flow strips on nitrocellulose membrane material and fabrication method. The 3D-printed POCT detector was used to supply a dark environment, with advantages of small volume, lightweight, and being easy-to-use. The smartphone-based data processing module was applied for image acquisition, image analysis and data display. After the fluorescent image was taken, the image analysis was carried out by the self-developed APP. By using the proposed POCT system, on the optimized conditions, the desired IgE detection was obtained, with a linear dynamic detection range of 10–1000 ng mL−1, and detection limit of 8.5 ng mL−1. In addition, the presented system showed good selectivity, repeatability and stability, and had the potential to be used in the field of detection of allergic diseases.

Development of an Allergic Rhinitis Diagnosis Application Using the Total Tear IgE Detection Kit for Examining Nasal Fluid: Comparison and Combination with the Conventional Nasal Smear Examination for Eosinophils

Allergic rhinitis (AR) is a type I allergic disease characterized by immunoglobulin E (IgE) -mediated hypersensitivity of the nasal mucosa. Here, we focused on a commercial test kit named Allerwatch® (AW) for the diagnosis of allergic conjunctivitis (AC) in which total tear IgE is qualitatively detected based on immunochromatography. We evaluated the usefulness of the AW test for detecting total IgE in the nasal discharge of AR and non-allergic rhinitis (non-AR) patients in comparison and combination with the conventional nasal smear examination for eosinophils. Using the AW test, total IgE in nasal fluid was detected in 64.76% of the AR patients and 11.11% of the non-AR patients, with a significant difference between the groups (p < 0.001). As compared to non-AR, the sensitivity and specificity of the detection of total IgE in nasal fluid for detecting AR were 64.76% and 88.89%, respectively. In the AR patients, house dust mites (57.1% of patients) and Japanese cedar pollen (93.3% of patients) were the major sensitizing antigens. When we considered a positive result in either of the two examinations to indicate a positive result, the rate of positivity in AR patients increased to 78.10%. As compared to non-AR, the sensitivity and specificity of the combination of both examinations for detecting AR were 78.10% and 83.33%, respectively. The AW test in the nasal cavity and the qualitative measurement of total IgE in nasal fluid may enable the detection of allergic elements in patients who present to a medical institution with nasal symptoms. In addition, the detection rate is increased when combined with the nasal smear examination for eosinophils.

The Humanised NPY-mRFP RBL Reporter Cell Line Is a Fast and Inexpensive Tool for Detection of Allergen-Specific IgE in Human Sera.

Rat basophilic leukaemia (RBL) cells have been used for decades as a model of high-affinity Immunoglobulin E (IgE) receptor (FcεRI) signalling. Here, we describe the generation and use of huNPY-mRFP, a new humanised fluorescent IgE reporter cell line. Fusion of Neuropeptide Y (NPY) with monomeric red fluorescent protein (mRFP) results in targeting of fluorescence to the granules and its fast release into the supernatant upon IgE-dependent stimulation. Following overnight sensitisation with serum, optimal release of fluorescence upon dose-dependent stimulation with allergen-containing extracts could be measured after 45 min, without cell lysis or addition of any reagents. Five substitutions (D194A, K212A, K216A, K226A, and K230A) were introduced into the FcεRIα cDNA used for transfection, which resulted in the removal of known endoplasmic reticulum retention signals and high surface expression of human FcεRIα* in huNPY-mRFP cells (where * denotes the penta-substituted variant), comparable to the ~500,000 FcεRIα molecules per cell in the RS-ATL8 humanised luciferase reporter, which is a human FcεRIα/FcεRIγ double transfectant. The huNPY-mRFP reporter was used to demonstrate engagement of specific IgE in sera of Echinococcus granulosus-infected individuals by E. granulosus elongation factor EgEF-1β and, to a lesser extent, by EgEF-1δ, which had been previously described as IgE-immunoreactive EgEF-1β/δ.

Immunological Changes of Basophil Hyperreactivity to Sweat in Patients With Well-Controlled Atopic Dermatitis.

BackgroundSweat aggravates atopic dermatitis (AD). In patients with AD, type-I hypersensitivity to sweat may be shown by histamine release from patients’ basophils in response to the semi-purified sweat antigen (QR), and the presence of specific immunoglobulin E (IgE) binding to MGL_1304, the component of QR. However, there has been no information on the immunological changes of type-I hypersensitivity to the sweat antigen in patients with well-controlled AD using topical corticosteroids (TCSs) and/or biologics as treatments.MethodHistamine-releasing tests using patients’ basophils and QR and the detection of serum IgE against MGL_1304 and mite allergen Der f 1 were performed in patients with AD who were well controlled by topical TCS with/without dupilumab for 53–96 weeks.ResultsIn total, 14 patients were enrolled. Seven patients received TCS therapy alone (TCS group), and seven patients received TCS with dupilumab therapy (dupilumab group). In all participants, the level of specific IgE against MGL_1304 decreased after treatments, but histamine release from basophils in response to QR did not show a statistically significant reduction; rather, it increased. In the dupilumab group, all changes in histamine release induced by QR (increase), the IgE level against MGL_1304 (decrease), and that against Der f 1 (decrease) were statistically significant, whereas the TCS group showed no significant change in any of them.ConclusionThe well-controlled condition for 53–96 weeks resulted in no reduction of the hyperreactivity of basophils against in patients with AD, even with the treatment with dupilumab. This study suggests persistent basophil hyperreactivity to sweat antigen over a year or longer.

Ultrasensitive detection of IgE levels based on magnetic nanocapturer linked immunosensor assay for early diagnosis of cancer

Rapid and accurate detection of immunoglobulin E (IgE) in serum and reduction of serum dosage are of great significance for clinical detection. Herein, we described a rapid magnetic separation of IgE from patient serum based on Fe3O4@SiO2-NTA@026 sdab as the capture probe and multiple horseradish peroxidase (HRP)-labeled antibodies linked gold nanoparticles (AuNPs) as chemiluminescence (CL) signal amplifier for ultrasensitive detection of total IgE. Results showed that the limit of detection of our immunosensor system in serum samples was 0.03 kU/L, which is lowest in comparison with current methods, and far lower than that of ImmunoCAP for IgE detection (0.1 kU/L). Furthermore, our immunosensor possessed satisfied repeatability and accuracy, as well as good stability. In comparison with the ImmunoCAP for the quantitative detection of IgE, highly consistent results were achieved in 20 serum samples. Specially, this method was also successfully utilized for assessing the IgE traces in breast cancer patients, which provides a new idea for the diagnosis of early cancer. Therefore, we believe that such versatile immunosensor will offer an alternative method for the on-site monitoring and determination of various IgE-related diseases.

Diagnostic value of combined detection of different indicators for type 2 diabetes mellitus combined with atherosclerotic disease

Objective: To investigate the diagnostic value of serum lipoprotein associated phospholipase A2 (Lp-PLA2), amyloid A (SAA) and immunoglobulin E (IgE) in patients with type 2 diabetes (T2DM) mellitus complicated with atherosclerotic disease. Methods: From June to December 2019, 224 patients with T2DM in the Second Hospital of Lanzhou University were selected, including 144 males and 80 females, aged (61±11) years. According to the results of imaging examination, the patients were divided into T2DM with AS group (T2DM-AS group, n=160) and T2DM group (n=64); Healthy subjects in the same period were selected as healthy control group (n=160). Lp-PLA2, IgE, SAA, hs-CRP, TC, TG, HDL-C, LDL-C and Hcy were detected in all patients and healthy controls. The correlation between the above indexes, gender, age and T2DM with AS was analyzed; Multivariate logistic regression was used to analyze the risk factors of T2DM with AS. Results: Compared with the healthy control group, the levels of IgE and Lp-PLA2 in T2DM-AS group and T2DM group were increased, and the levels of SAA in T2DM-AS group were increased (all P<0.05); Compared with T2DM group, the levels of Lp-PLA2, IgE and SAA were increased in T2DM-AS group (all P<0.05). T2DM with AS was positively correlated with age, IgE, Lp-PLA2 and SAA (r=0.468, 0.269, 0.486, 0.418, all P<0.05), and negatively correlated with HDL-C (r=-0.338, P<0.05). Multivariate logistic regression analysis showed that age (OR=0.865, 95%CI: 0.763-0.982, P<0.05), IgE (OR=0.910, 95%CI: 0.840-0.987, P<0.05) and Lp-PLA2 (OR=0.942, 95%CI: 0.910-0.986, P<0.05) were risk factors of T2DM with AS. ROC curve showed that the combined detection of Lp-PLA2, SAA and IgE could improve the diagnostic efficiency of T2DM with AS (AUC=0.895, P<0.05), the sensitivity was 80.0%, and the specificity was 93.7%. Conclusion: The levels of Lp-PLA2, IgE and SAA increase in T2DM patients with AS. The combined detection of Lp-PLA2, SAA and IgE can improve the diagnostic efficiency of T2DM patients with AS.

Serum IgE Predicts Difference of Population and Allergens in Allergic Diseases: Data from Weifang City, China.

Background Immunoglobulin E (IgE) is the most important promoter of allergic inflammation. However, there are few systematic studies on IgE in age range, genders, disease spectrum, and time regularity. Aim To screen the common allergens, allergen spectrum, and IgE difference between type 2 inflammatory allergic diseases and other allergic diseases in Weifang, China. Methods A retrospective study was performed by estimating patients' clinical data suffering from allergic diseases (urticaria, pollinosis, allergic rhinitis, atopic dermatitis, and bronchial asthma) between May 2019 and April 2020 using an allergen detection kit of Macro-Union Pharmaceutical. Results 732 of the 1367 patients showed different antigen positive, and the positive rate was 53.5%. The most common allergens were dust mites, mixed fungi, Artemisia pollen, cat/dog dander, and cockroaches. There were 27.0% (369/1367) of the patients with single positive allergen-specific IgE (sIgE), 26.5% (363/1367) with multiple-positive IgE. The total immunoglobulin E (tIgE) levels varied with gender, age, and type of disease. There was a difference in the distribution of allergens between children and adults. A positive correlation between the serum-specific IgE and the corresponding local inhaled allergen density was observed. Conclusions In this study, we found that type 2 inflammatory allergic diseases have higher serum IgE and a higher probability of inhaled sIgE positive. According to age, gender, and condition, serological IgE detection of allergens provides new insight into the early diagnosis and prevention of allergic diseases.

Microarray-Based Allergy Diagnosis: Quo Vadis?

More than 30% of the world population suffers from allergy. Allergic individuals are characterized by the production of immunoglobulin E (IgE) antibodies against innocuous environmental allergens. Upon allergen recognition IgE mediates allergen-specific immediate and late-phase allergic inflammation in different organs. The identification of the disease-causing allergens by demonstrating the presence of allergen-specific IgE is the key to precision medicine in allergy because it allows tailoring different forms of prevention and treatment according to the sensitization profiles of individual allergic patients. More than 30 years ago molecular cloning started to accelerate the identification of the disease-causing allergen molecules and enabled their production as recombinant molecules. Based on recombinant allergen molecules, molecular allergy diagnosis was introduced into clinical practice and allowed dissecting the molecular sensitization profiles of allergic patients. In 2002 it was demonstrated that microarray technology allows assembling large numbers of allergen molecules on chips for the rapid serological testing of IgE sensitizations with small volumes of serum. Since then microarrayed allergens have revolutionized research and diagnosis in allergy, but several unmet needs remain. Here we show that detection of IgE- and IgG-reactivity to a panel of respiratory allergens microarrayed onto silicon elements is more sensitive than glass-based chips. We discuss the advantages of silicon-based allergen microarrays and how this technology will allow addressing hitherto unmet needs in microarray-based allergy diagnosis. Importantly, it described how the assembly of silicon microarray elements may create different microarray formats for suiting different diagnostic applications such as quick testing of single patients, medium scale testing and fully automated large scale testing.

Paper-based upconversion fluorescence aptasensor for the quantitative detection of immunoglobulin E in human serum

Immunoglobulin E (IgE), a biomarker of allergic diseases, plays a critical role in allergic mechanism. Because of its low abundance in serum, the demand of developing sensitive, selective and simple methods for IgE detection is still very urgent. Paper-based analytical devices using upconversion nanoparticles (UCNPs) as the label can be promising point-of-care test (POCT) methods in rapid diagnosis, owing to their NIR-excitation and visible light emission nature, which can avoid the interference of autofluorescence and scattering light from biological samples and paper substrates. In this work, we proposed a paper-based analytical device for the sensitive, selective and accurate detection of total immunoglobulin E (IgE) in human serum. The assay was based on resonance energy transfer between UCNPs and organic dye tetramethylrhodamine (TAMRA), and IgE aptamer with stem-loop structure was used as the recognizing probe. The existence of IgE change the conformation of IgE aptamer, enlarge the distance between donor and acceptor, and block the energy transfer process. Thus, the luminescence of UCNPs recovered with an IgE concentration independent manner. A linear calibration was obtained in the range of 0.5–50 IU/mL, with a detection limit of 0.13 IU/mL. The results of our method were well correlated with that of commercial ELISA kit (20 human serum samples). This work suggests promising prospect of the paper-based UC-LRET analytical devices in real samples and may promote the application of paper-based analytical devices in clinical diagnosis.

Galactose-alpha-1,3-galactose syndrome.

The galactose-alpha-1,3-galactose (alpha-Gal) syndrome is a newly recognized and unique form of food allergy, characterized by delayed reactions to mammalian meats. This form of allergy occurs in individuals who become sensitized to alpha-Gal, a carbohydrate that is present on most mammalian tissues. Sensitization occurs after exposure to multiple arthropod bites, most commonly the lone star tick. Cases of the alpha-Gal syndrome are primarily found in the southeastern United States, which overlaps with the known geographic distribution of the lone star tick. Patients present with a history of delayed symptom onset, ~2-6 hours after ingestion of mammalian meat. As with other immunoglobulin E (IgE) mediated food allergic reactions, alpha-Gal reaction symptoms may include skin, respiratory, gastrointestinal, or cardiovascular systems, and severity may range from mild reactions to severe anaphylaxis. The diagnosis is based on the detection of alpha-Gal specific IgE (sIgE) as well as the total IgE value because some cases include patients with low total IgE levels but a high percentage of alpha-Gal sIgE to total serum IgE levels. Percutaneous testing with commercial meat skin-prick testing extracts is not a reliable tool for diagnosis. Prick-prick skin testing to fresh cooked meat may be considered, whereas intradermal testing to fresh meat is primarily reserved for research purposes. The mainstay of treatment involves avoidance of mammalian meat and medications that express the same carbohydrate antigen. With a small portion of patients, other meat-containing products should also be avoided if symptoms persist with mammalian meat avoidance alone. Prolonged avoidance of mammalian meat as well as avoidance of further tick bites can decrease alpha-Gal sIgE over time, and some patients are able to reintroduce mammalian meat into their diet.

Microarray-Based Detection of Allergen-Reactive IgE in Patients with Mastocytosis

Because of a high risk to develop fatal anaphylaxis, early detection of immunoglobulin E (IgE)-dependent allergy is of particular importance in patients with mastocytosis. We examined whether microarray-based screening for allergen-reactive IgE (allergen-chip) is a sensitive and robust approach to detect specific IgE in patients with mastocytosis. Sera for 42 patients were analyzed, including 4 with cutaneous mastocytosis, 2 with mastocytosis in the skin, and 36 with systemic mastocytosis. In addition, sera from an age- and sex-matched control cohort (n= 42) were analyzed. In 15 of 42 patients with mastocytosis (35.7%), specific IgE was detected by allergen-chip profiling. Ves v 5 and Bet v 1 were the most frequently detected allergens (Ves v 5: 16.7% of patients; Bet v 1: 11.9% of patients). Allergen reactivity was confirmed by demonstrating upregulation of CD203c on blood basophils upon exposure to the respective allergen(s) in these patients. Specific IgE was identified by chip studies in 11 of 26 patients with mastocytosis with mediator-related symptoms (42.3%) and in 4 of 14 patients with mastocytosis without symptoms (28.6%). In the cohort with known allergy, 9 of 9 patients (100%) had a positive allergen-chip result. In patients with mastocytosis without a known allergy (n= 31), the chip identified 6 positive cases (19.5%). The prevalence of chip-positive patients was slightly lower in the mastocytosis group (35.7%) compared with age- and sex-matched controls (40.5%). Although specific IgE may not be detectable in all sensitized patients with mastocytosis, allergy chip-profiling is a reliable screening approach for the identification of patients with mastocytosis suffering from IgE-dependent allergies.

Electrochemical Immunosensor for Human IgE Using Ferrocene Self-Assembled Monolayers Modified ITO Electrode.

The immunoglobulin E (IgE) level in serum is an important factor in the examination of allergy. Ferrocene (Fc)-modified self-assembled monolayers (SAMs) were placed on an indium tin oxide (ITO) electrode as a sensing layer for the detection of human IgE. The Fc moiety in the SAMs facilitated the electron transfer through the organic SAMs layer and electrocatalytic signal amplification. The electrochemical measurement was accomplished after the sandwich type immobilization of the receptor antibody, target human IgE, and enzyme conjugated secondary antibody. The enzyme product, p-aminophenol, was quantitatively analyzed by redox cycling via Fc. In addition, the electrochemical impedance spectroscopy (EIS) was investigated for the detection of IgE. The limit of detection (LOD), limit of quantification (LOQ), and dynamic range of the electrochemical sensor were 3 IU/mL, 10 IU/mL, and from 10 IU/mL to 100 IU/mL, respectively.

Directing a rational design of aptamer-based fluorescence anisotropy assay for sensitive detection of immunoglobulin E by site-specific binding study

Mapping aptamer-protein interactions is important for characterization and applications of aptamers against proteins. We describe here probing affinity interactions between aptamer and immunoglobulin E (IgE) with a fluorescence anisotropy (FA) approach using a series of aptamer probes having single fluorescein (FAM) label at individual nucleotide (A, C, T). Studies of binding between IgE and aptamer probes revealed several possible close-contact sites, e.g., T9, T10, T11, T13, C15, and T17 of a 37-nt aptamer with a stem-loop secondary structure. FAM labeling on these sites resulted in much higher FA values (higher than 0.230 for T10, T11, T13 and C15) of aptamer-IgE complexes than the distant sites (e.g., terminals) of aptamer probably because the bound IgE close to these sites significantly restricted local rotation of FAM. Close-contact site labeled aptamer probes with high affinity allowed to develop a more sensitive FA assay for IgE than distant site labeled aptamers. The FA assay using T10-labeled aptamer with a dissociation constant (Kd) about 0.8 nM enabled selective detection of IgE at 20 pM and large FA increase upon IgE addition. We also found A12, C14, A25, and T27 were important for IgE-aptamer binding as FAM labeling at these sites significantly reduced aptamer affinity. FA study showed the loop region of this stem-loop aptamer was crucial for affinity binding, and IgE bound to the loop. This FA method will be helpful for understanding aptamer-protein binding and making a rational design of aptamer affinity assays for proteins.

A quantum dot-based lateral flow immunoassay for the rapid, quantitative, and sensitive detection of specific IgE for mite allergens in sera from patients with allergic rhinitis

The prevalence of allergic rhinitis (AR) is increasing worldwide. However, the current systems used to measure levels of immunoglobulin E (IgE) in sera are associated with several disadvantages that limit their further application. Consequently, there is a need to develop novel highly sensitive strategies that can rapidly detect IgE in a quantitative manner. The development of such systems will significantly enhance our ability to diagnose, treat, and even prevent AR. Herein, we describe our experience of using quantum dot-based lateral flow immunoassay (QD-LFIA), combined with a portable fluorescence immunoassay chip detector (PFICD), to detect serum-specific IgE against Dermatophagoides pteronyssinus (Der-p) and Dermatophagoides farinae (Der-f), two common mite allergens in China. Our data showed that our system could detect serum-specific levels of IgE against Der-p and Der-f as low as 0.093 IU/mL and 0.087 IU/mL, respectively. We also established a standard curve to determine serum-specific IgE concentrations that correlated well with the clinical BioIC microfluidics system. The sensitivity of our assay was 96.7% for Der-p and 95.5% for Der-f, while the specificity was 87.2% for Der-p and 85.3% for Der-f. Collectively, our results demonstrate that QD-LFIA is a reliable system that could be applied to detect serum-specific IgE in accordance with clinical demands. This QD-LFIA strategy can be applied at home, in hospitals, and in pharmacies, with reduced costs and time requirements when compared with existing techniques. In the future, this system could be developed to detect other types of allergens and in different types of samples (for example, whole blood).Graphical abstractWe describe our experiment using a quantum dot-based lateral flow immunoassay combined with a portable fluorescence immunoassay chip detector for both qualitative and quantitative detection of serum-specific IgE against two common mite allergens. This strategy can be applied at home, in hospitals, and in pharmacies, with reduced costs and time requirements. In the future, this system could be developed to detect other types of allergens and in different types of samples.

Antigen array for serological diagnosis and novel allergen identification in severe equine asthma

Severe equine asthma (sEA), which closely resembles human asthma, is a debilitating and performance-limiting allergic respiratory disorder which affects 14% of horses in the Northern Hemisphere and is associated with increased allergen-specific immunoglobulin E (IgE) against a range of environmental proteins. A comprehensive microarray platform was developed to enable the simultaneous detection of allergen-specific equine IgE in serum against a wide range of putative allergenic proteins. The microarray revealed a plethora of novel pollen, bacteria, mould and arthropod proteins significant in the aetiology of sEA. Moreover, the analyses revealed an association between sEA-affected horses and IgE antibodies specific for proteins derived from latex, which has traditionally been ubiquitous to the horse’s environment in the form of riding surfaces and race tracks. Further work is required to establish the involvement of latex proteins in sEA as a potential risk factor. This work demonstrates a novel and rapid approach to sEA diagnosis, providing a platform for tailored management and the development of allergen-specific immunotherapy.

Self-Assembly of Functional Nucleic Acid-Based Colorimetric Competition Assay for the Detection of Immunoglobulin E.

In this work, we have developed a simple and rapid colorimetric assay for the detection of immunoglobulin E (IgE) using functional nucleic acids (FNAs) and a solid-phase competition enzyme-linked immunosorbent assay (ELISA). The FNAs including aptamer of recombinant IgE, G-quadruplex and its complementary fragments were immobilized on 96-well microplates to achieve recognition and detection of IgE in biological samples. The G-quadruplex DNAzyme catalyzed 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS)-hemin-H2O2 system was used to improve the sensitivity of colorimetric assay. In the presence of IgE, the hairpin structure and G-quadruplex would be destroyed, resulting in the inactivation of DNAzyme and subsequent reduction of its absorbance. This cost-effective approach detected IgE in the linear range from 5.0 pg/mL to 500 ng/mL, with the limit of detection (LOD) of 2.0 pg/mL, under optimal conditions. Moreover, the developed method was successfully applied to the rapid detection of IgE in human urine, indicating a great potentiality of this approach in clinical diagnosis and other biomedical applications.

Detection of IgE, Some Antioxidant Parameter Levels and their Association with Polymorphism of ADAM33 Gene in Asthma Patients

Asthma is a multifactorial disease influenced by interactions between genetic and environmental factors. The alterations in some immunological, genetic and antioxidant markers can affecting pathogenesis of asthma, therefore, this study aimed to assessment of a disintegrin and metalloproteinase 33 V4 G/C (ADAM33 V4 C/G) polymorphism and its associated immunoglobulin E (IgE) antibodies and some antioxidant parameters with asthma in some Iraqi asthmatic patients. Blood samples collected from seventy asthmatic patients and forty healthy subjects (males and females) aged (18-70 year). Enzyme linked immunosorbent assay has been used for serum IgE estimation and other antioxidant parameters were carry out using biochemical methods for both groups. Genomic DNA was extracted from all blood samples and detected of ADAM33 V4 C/G polymorphism using Polymerase Chain Reaction – restriction fragment length polymorphism (PCR-RFLP) technique. Results showed that : there was a significant increase in IgE concentration (p 0.01) in asthmatic patients compared with controls. there was a highly significant differences in alleles and genotypes frequency between patient and healthy group (p< 0.0001), frequency of G allele and genotype GG were higher in asthmatic patients compared with controls and there was a correlation between high IgE level and the presence of allele G within the patient genotypes. We can conclude that concentration of IgE and frequency of GG genotype were elevated in asthmatic patients compared with healthy. GG genotype and allele G can be considered as a genetic marker of the disease and presence of this allele is associated with the high levels of IgE and increase of disease severity.

Study of the role of allergy diagnosed by immunoglobulin E in the etiology of pediatric otitis media with effusion

Objectives This study aimed to study the role of allergy diagnosed by immunoglobulin E (IgE) in the etiology of pediatric otitis media with effusion (OME) by detection of IgE in middle-ear effusion using the enzyme-linked immunosorbent assay technique. Background The role of allergy in chronic OME is controversial. Patients and methods This was a prospective study that was carried out on 80 cases; including 60 pediatric patients diagnosed with chronic OME [resistant to medical treatment for >3 months (type B tympanogram) or recurrent after successful medical treatment]. Patients were subjected to a myringotomy with insertion of a tympanostomy tube (grommet tube). Their age ranged from 2 to 15 years. Twenty children had ENT disease other than OME, mostly adenoidal or tonsillar hypertrophy. The serum level of total IgE was assayed in patients and control groups as well as in the middle-ear effusion of patients. Results The results showed that total IgE was higher in the serum of patients with OME than the serum of the control group; there was a high significant correlation between total IgE in serum and total IgE in effusion in patients with OME. Conclusion Allergy is a possible risk factor for the development of pediatric OME.