The Western blot is a highly sensitive and efficient technique in diagnosing allergy to wasp venom

Zollner, Thomas M.; Spengler, K.; Ergezinger, Katrin; Kaufmann, Roland; Boehncke, Wolf‐Henning

doi:10.1046/j.1365-2222.2001.01210.x

Cited by 18 publications

(15 citation statements)

References 26 publications

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“…Other in vitro tests When venom skin tests and the measurement of venom‐specific IgE antibodies in serum by RAST or an equivalent method yield negative results in patients with a history of a systemic anaphylactic sting reaction, additional in vitro tests may be used to demonstrate immunologic sensitization (like immunoblotting, the basophil histamine release test, basophil activation test and leukotrine release test) (122–126).…”

mentioning

confidence: 99%

Diagnosis of Hymenoptera venom allergy

Bilò¹,

Ruëff

Mosbech³

et al. 2005

Allergy

503

439

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The first EAACI position paper on immunotherapy with Hymenoptera venoms was published in 1987 (1). Six years later a revised version appeared (2). As then many papers on the diagnosis and treatment of Hymenoptera venom allergy have been published, making a review of the last EAACI position paper necessary. This paper focuses on Hymenoptera venom allergy, as allergic reactions caused by stings of insects other than Hymenoptera are rare and standardized extracts for the diagnosis and treatment of allergic reactions to nonHymenoptera insects are not available (3). TaxonomyMost authors follow the Chinery classification (4), although over the last few years a few minor changes have been introduced. Aculeatae are a suborder of Hymenoptera (Fig. 1).The family Apidae consists of the honeybees (Apis mellifera) who are brown in color and moderately hairy and the bumblebees (Genus Bombus) which are bigger than honeybees, much more hairy and characterized by distinct yellow or white bands on their abdomen. Vespidae are divided into the Vespinae and Polistinae subfamilies, with differences at the junction of the thorax and abdomen. Vespinae have a truncated junction while Polistinae are more oval in shape. Vespidae are almost hairless and have black and yellow striped abdomens.Vespula, Dolichovespula and Vespa make up the three genera of the Vespinae. Vespula (called wasps in Europe, yellow jackets in the USA) are the most important species in Europe. The Vespula spp. (V. germanica and The purpose of diagnostic procedure is to classify a sting reaction by history, identify the underlying pathogenetic mechanism, and identify the offending insect. Diagnosis of Hymenoptera venom allergy thus forms the basis for the treatment. In the central and northern Europe vespid (mainly Vespula spp.) and honeybee stings are the most prevalent, whereas in the Mediterranean area stings from Polistes and Vespula are more frequent than honeybee stings; bumblebee stings are rare throughout Europe and more of an occupational hazard. Several major allergens, usually glycoproteins with a molecular weight of 10-50 kDa, have been identified in venoms of bees, vespids. and ants. The sequences and structures of the majority of venom allergens have been determined and several have been expressed in recombinant form. A particular problem in the field of cross-reactivity are specific immunoglobulin E (IgE) antibodies directed against carbohydrate epitopes, which may induce multiple positive test results (skin test, in vitro tests) of still unknown clinical significance. Venom hypersensitivity may be mediated by immunologic mechanisms (IgE-mediated or non-IgE-mediated venom allergy) but also by nonimmunologic mechanisms. Reactions to Hymenoptera stings are classified into normal local reactions, large local reactions, systemic toxic reactions, systemic anaphylactic reactions, and unusual reactions. For most venom-allergic patients an anaphylactic reaction after a sting is very traumatic event, resulting in an altered health-related quality of life. Risk f...

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mentioning

confidence: 99%

Diagnosis of Hymenoptera venom allergy

Bilò¹,

Ruëff

Mosbech³

et al. 2005

Allergy

503

439

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“…In particular, the usefulness of the cellular assays CAST-ELISA and Flow-CAST in the management of the HV allergy was investigated respectively in 3 and 5 patient control studies selected in this review [1][2][3][16][17][18]. In the CAST-ELISA studies were included 35 subjects with negative responses for at least one of the standard tests, and 2 additional subjects with negative results for both usual methods (see the Table 1).…”

Section: Resultsmentioning

confidence: 99%

“…A further in vitro technique recently used in the diagnosis of HV allergy is the Western blot [18]. In this method allergen extracts consist of a complex mixture of proteins which can be separated by molecular size on SDS-PAGE.…”

Section: Resultsmentioning

confidence: 99%

Approaches to the Management of Presumed Immediate Hymenoptera Venom Allergy and Non-Detectable IgE

Mingomataj¹,

Bakiri²

2010

TOALLJ

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Objective: To provide a comprehensive evaluation in patients with a convincing history of immediate insect allergy but negative skin test and/or specific IgE results, adequately addressing the question of how best to manage them. Data sources: Observational peer-reviewed studies and case reports were searched on Pub-Med database from 1998 up to March 2009 using the following keywords: Hymenoptera Allergy & Negative IgE (Negative Skin Tests). Study selection: Studies on supplemental diagnostic tests that provided data from patients with immediate hymenoptera allergy but negative conventional tests results to the offending allergens were selected. In this work, we also included studies providing additional relevant information regarding this issue. Results: Among 43 identified papers only 9 of them presented relevant original data, while the other papers were reviews. In the majority of the cases, the culprit insect was identified with in vitro tests such as Basophil Activation Test, Cellular Allergen Stimulation Test or Western blot, whereas in vivo (less frequently) with sting challenge or dialyzed venom skin test. Conclusions: The management of patients with a convincing history of immediate insect allergy but negative conventional test results requires an adaption of the guidelines including an incorporation of the novel diagnostic tools. Although cellular tests represent equivalent sensitivity and superior specificity as compared with standard ones, these tests still remain supplementary diagnostic tools. In a minority of cases (especially in the developing countries where cellular tests cannot be performed), venom immunotherapy in adult subjects could be taken into account based solemnly on the history of a clear patient’s identification of the culprit insect.

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“…As previously reported, Western blot is a highly sensitive and efficient technique in detecting specific IgE [23-25]. C. pneumoniae (TW183) and C. trachomatis (serovar E) elementary bodies were purified by 20%–50% (vol/vol) Renografin gradient centrifugation as previously described [26] and normalized for protein content using the Bradford protein assay.…”

Section: Methodsmentioning

confidence: 99%

The prevalence and identity of Chlamydia -specific IgE in children with asthma and other chronic respiratory symptoms

et al. 2012

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BackgroundRecent studies have confirmed the presence of viable Chlamydia in the bronchoalveolar lavage (BAL) fluid of pediatric patients with airway hyperresponsiveness. While specific IgG and IgM responses to C. pneumoniae are well described, the response and potential contribution of Ag-specific IgE are not known. The current study sought to determine if infection with Chlamydia triggers the production of pathogen-specific IgE in children with chronic respiratory diseases which might contribute to inflammation and pathology.MethodsWe obtained BAL fluid and serum from pediatric respiratory disease patients who were generally unresponsive to corticosteroid treatment as well as sera from age-matched control patients who saw their doctor for wellness checkups. Chlamydia-specific IgE was isolated from BAL and serum samples and their specificity determined by Western blot techniques. The presence of Chlamydia was confirmed by species-specific PCR and BAL culture assays.ResultsChlamydial DNA was detected in the BAL fluid of 134/197 (68%) patients. Total IgE increased with age until 15 years old and then decreased. Chlamydia-specific IgE was detected in the serum and/or BAL of 107/197 (54%) patients suffering from chronic respiratory disease, but in none of the 35 healthy control sera (p < 0.0001). Of the 74 BAL culture-positive patients, 68 (91.9%, p = 0.0001) tested positive for Chlamydia-specific IgE. Asthmatic patients had significantly higher IgE levels compared to non-asthmatics (p = 0.0001). Patients who were positive for Chlamydia DNA or culture had significantly higher levels of serum IgE compared to negative patients (p = 0.0071 and p = 0.0001 respectively). Only 6 chlamydial antigens induced Chlamydia-specific IgE and patients with C. pneumoniae-specific IgE had significantly greater levels of total IgE compared to C. pneumoniae-specific IgE negative ones (p = 0.0001).ConclusionsIgE antibodies play a central role in allergic inflammation; therefore production of Chlamydia-specific IgE may prove significant in the exacerbation of chronic, allergic airway diseases, thus highlighting a direct role for Chlamydia in asthma pathogenesis.

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The Western blot is a highly sensitive and efficient technique in diagnosing allergy to wasp venom

Cited by 18 publications

References 26 publications

Diagnosis of Hymenoptera venom allergy

Diagnosis of Hymenoptera venom allergy

Approaches to the Management of Presumed Immediate Hymenoptera Venom Allergy and Non-Detectable IgE

The prevalence and identity of Chlamydia -specific IgE in children with asthma and other chronic respiratory symptoms

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