The major Alternaria alternata allergen, Alt a 1: A reliable and specific marker of fungal contamination in citrus fruits.

The ubiquitously present spores of Alternaria alternata can spoil a wide variety of foodstuffs, including a variety of fruits belonging to the Citrus genus. The major allergenic protein of A. alternata, Alt a 1, is a species-specific molecular marker that has been strongly associated with allergenicity and phytopathogenicity of this fungal species. This study aimed to evaluate the potential of the detection of Alt a 1 as a reliable indicator of A. alternata contamination in citrus fruits. To accomplish this aim, sixty oranges were artificially infected with a spore suspension of A. alternata. Internal fruit material was collected at different incubation times (one, two and three weeks after the fungal inoculation) and used for both total RNA extraction and protein extraction. Alt a 1 detection was then performed by polymerase chain reaction (PCR) amplification using Alt a 1 specific primers and by enzyme-linked immunosorbent assay (ELISA). The experimental model presented in this work was effective to simulate the typical Alternaria black rot phenotype and its progression. Although both PCR and ELISA techniques have been successfully carried out for detecting Alt a 1 allergen in A. alternata infected oranges, the PCR method was found to be more sensitive than ELISA. Nevertheless, ELISA results were highly valuable to demonstrate that considerable amounts of Alt a 1 are produced during A. alternata fruit infection process, corroborating the recently proposed hypothesis that this protein plays a role in the pathogenicity and virulence of Alternaria species. Such evidence suggests that the detection of Alt a 1 by PCR-based assay may be used as a specific indicator of the presence of pathogenic and allergenic fungal species, A. alternata, in fruits. This knowledge can be employed to control the fungal infection and mitigate agricultural losses as well as human exposure to A. alternata allergens and toxins.

IgE-binding aldolases from artemia and mysis species causing respiratory allergy to aquarium fishfood

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Alt a 15 is a new cross-reactive minor allergen of Alternaria alternata.

Alternaria alternata is one of the most common saprophytes worldwide that is clinically and epidemiologically associated with severe asthma. Therefore, the identification and characterization of all A. alternata allergens are of major clinical importance. This study describes a new cross-reactive A. alternata allergen that was officially named Alt a 15 by the official Allergen Nomenclature Subcommittee. The complete coding region for Alt a 15 was amplified using 5' and 3' rapid amplification of cDNA ends and PCR. The recombinant protein was produced in Escherichia coli as a 65-kDa fusion protein, and the protein sequence exhibits high homology with several important fungal allergens. Immunoblotting analyses revealed that IgE antibodies from A. alternata-sensitized patients (n=59) bound to rAlt a 15 with a prevalence of 10.2%. All patients who presented sIgE to rAlt a 15 were apparently poly-sensitized to A. alternata and C. lunata. The extensive cross-reactivity between A. alternata and C. lunata serine proteases was confirmed using immunoblotting inhibition assays. Overall, Alt a 15 is an important new cross-reactive allergen of A. alternata that explains some allergies to A. alternata without Alt a 1 sensitization and initial diagnostic errors for allergies to Alternaria. This molecule may improve the accuracy of the diagnosis, the understanding, and the management of IgE-mediated fungal diseases.

Development of a PCR-based tool for detecting immunologically relevant Alt a 1 and Alt a 1 homologue coding sequences.

Alt a 1 has been recognized as the most important allergen produced by the Pleosporaceae family and is a risk factor for asthma development and/or exacerbation. The aim of this study was to develop a detection tool that is highly specific for species that produced airborne Alt a 1. Based on the highly conserved internal nucleotide region of the several Alt a 1 sequences that are available in GenBank, a pair of primers (Alta1CF/Alta1CR) was designed. A set of primers used by other authors for the production of recombinant Alt a 1 (A21F/A21R) was also tested. The molecular analyses were based on the polymerase chain reaction (PCR) amplification and sequencing of the cDNA that was obtained from thirteen common fungal species. The PCR system that utilized Alta1CF/Alta1CR was highly specific, sensitive, and was able to detect an amplicon of approximately 180 bp from Alt a 1 and Alt a 1-like encoding genes from A. alternata, A. tenuissima, A. infectoria, U. botrytis, and S. botryosum. In contrast, the A21F/A21R primers were specific for the very close taxonomically related species A. alternata and A. tenuissima. Thus, this rapid, sensitive, and specific detection tool can be used to assess Alt a 1 exposure levels and to inform the implementation of the appropriate public health measures.

Evaluation and Comparison of Commercially Available Latex Extracts for Skin Prick Tests

Background: Crude latex extracts are commonly used in skin prick tests (SPT) for the diagnosis of natural rubber latex (NRL) allergy. Nevertheless, variations in protein and allergen composition between latex extracts from different manufacturers can hamper a correct diagnosis. Objectives: To analyze the heterogeneity of proteins and allergens in latex extracts from 7 different manufacturers and to assess its relevance in the diagnosis of latex allergy. Methods: Seven latex SPT extracts were analyzed for protein content using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDSPAGE). The 4 major allergens Hev b 1, Hev b 3, Hev b 5, and Hev b 6.02 were also quantified using enzyme immunoassay. All commercial extracts were tested for their in vitro allergenic capacity using microarray inhibition assays and for their ability to induce biological reactivity in latex-allergic patients undergoing SPT. Results: The protein content of the extracts varied widely from 8.0 μg/mL to 526.5 μg/mL. SDS-PAGE revealed broad differences in protein profiles between the extracts. Marked variability in the contents of all 4 major allergens was observed, and Hev b 3 and Hev b 5 were undetectable in some extracts. Microarray inhibition assays and SPT demonstrated relevant differences in allergenic capacity between the extracts. Conclusions: The marked heterogeneity in protein and allergen content of latex extracts from different manufacturers could explain the broad spectrum of SPT results recorded. Our fi ndings suggest that the extracts used for the diagnosis of latex allergy should be improved and standardized (AU)

Antecedentes: En el diagnóstico de la alergia a látex natural se utilizan habitualmente extractos crudos de látex en técnica de puntura. No obstante la variación que existe en el contenido proteico y de los distintos alérgenos entre los extractos comerciales procedentes de distintos fabricantes podría afectar al correcto diagnóstico de la alergia. Objetivos: Analizar la heterogeneidad proteica y de alérgenos entre siete extractos de látex de distintos fabricantes y comprobar las posibles implicaciones clínicas en el diagnóstico de la alergia al látex. Métodos: Se analizó el contenido proteico de siete extractos de látex y también el perfil mediante la técnica de electroforesis en gel de poliacrilamida (SDS-PAGE). Además, se cuantificaron mediante enzimo-inmunoensayo (EIA), los cuatro alérgenos principales de látex Hev b 1, Hev b 3, Hev b 5 y Hev b 6.02. También se estudió en los siete extractos comerciales su capacidad de inhibición "in vitro", del ensayo de micromatrices y su capacidad para inducir respuestas biológicas "in vivo" en pacientes con alergia al látex, mediante pruebas cutáneas en puntura (SPT). Resultados: Los extractos presentaban una amplia variación en el contenido proteico que oscilaba entre 8.0 y 526.5 μg/mL de extracto. También se observaron importantes diferencias en el perfil proteico mediante la técnica de SDS-PAGE. El contenido de los principales cuatro alérgenos fue también muy variable, de forma que en algunos extractos los contenidos de Hev b 3 y Hev b 5 fueron prácticamente indetectables. Tanto la técnica de inhibición de micromatrices como las pruebas de puntura mostraron diferencias notables en la capacidad alergénica de los distintos extractos. Conclusiones: Los extractos de látex provenientes de distintos fabricantes presentan una importante heterogeneidad en contenido proteico y de alérgenos que podría claramente explicar las notables diferencias observadas en los resultados de las pruebas cutáneas en puntura que presentan los pacientes. Nuestros resultados apoyan la necesidad de mejora de la estandarización de los extractos de látex habitualmente utilizados en el diagnóstico clínico de la alergia al látex (AU)

Clinical value of morphine, pholcodine and poppy seed IgE assays in drug-abusers and allergic people

Background: The diagnosis of anaphylactic reactions due to opiates during anaesthesia can be difficult, since in most cases various drugs may have been administered. Detection of specific IgE to poppy seed might be a marker for sensitisation to opiates in allergic people and heroin-abusers. This study assessed the clinical value of morphine, pholcodine and poppy seed skin-prick and IgE determination in people suffering hypersensitivity reactions during anaesthesia or analgesia and drug-abusers with allergic symptoms. Methods: We selected heroin abusers and patients who suffered severe reactions during anaesthesia and analgesia from a database of 23,873 patients. The diagnostic yield (sensitivity, specificity and predictive value) of prick and IgE tests in determining opiate allergy was analysed. Results: Overall, 149 patients and 200 controls, mean age 32.9±14.7 years, were included. All patients with positive prick to opiates showed positive prick and IgE to poppy seeds, but not to morphine or pholcodine IgE. Among drug-abusers, 13/42 patients (31%) presented opium hypersensitivity confirmed by challenge tests. Among non-drug abusers, sensitisation to opiates was higher in people allergic to tobacco (25%), P<0.001. Prick tests and IgE against poppy seed had a good sensitivity (95.6% and 82.6%, respectively) and specificity (98.5% and 100%, respectively) in the diagnosis of opiate allergy. Conclusions: Opiates may be significant allergens. Drug-abusers and people sensitised to tobacco are at risk. Both the prick and specific IgE tests efficiently detected sensitisation to opiates. The highest levels were related to more-severe clinical profiles(AU)

Clinical value of morphine, pholcodine and poppy seed IgE assays in drug-abusers and allergic people.

BACKGROUND: The diagnosis of anaphylactic reactions due to opiates during anaesthesia can be difficult, since in most cases various drugs may have been administered. Detection of specific IgE to poppy seed might be a marker for sensitisation to opiates in allergic people and heroin-abusers. This study assessed the clinical value of morphine, pholcodine and poppy seed skin-prick and IgE determination in people suffering hypersensitivity reactions during anaesthesia or analgesia and drug-abusers with allergic symptoms. METHODS: We selected heroin abusers and patients who suffered severe reactions during anaesthesia and analgesia from a database of 23,873 patients. The diagnostic yield (sensitivity, specificity and predictive value) of prick and IgE tests in determining opiate allergy was analysed. RESULTS: Overall, 149 patients and 200 controls, mean age 32.9 ± 14.7 years, were included. All patients with positive prick to opiates showed positive prick and IgE to poppy seeds, but not to morphine or pholcodine IgE. Among drug-abusers, 13/42 patients (31%) presented opium hypersensitivity confirmed by challenge tests. Among non-drug abusers, sensitisation to opiates was higher in people allergic to tobacco (25%), P<.001. Prick tests and IgE against poppy seed had a good sensitivity (95.6% and 82.6%, respectively) and specificity (98.5% and 100%, respectively) in the diagnosis of opiate allergy. CONCLUSIONS: Opiates may be significant allergens. Drug-abusers and people sensitised to tobacco are at risk. Both the prick and specific IgE tests efficiently detected sensitisation to opiates. The highest levels were related to more-severe clinical profiles.

Evaluation and comparison of commercially available latex extracts for skin prick tests.

BACKGROUND: Crude latex extracts are commonly used in skin prick tests (SPT) for the diagnosis of natural rubber latex (NRL) allergy. Nevertheless, variations in protein and allergen composition between latex extracts from different manufacturers can hamper a correct diagnosis. OBJECTIVES: To analyze the heterogeneity of proteins and allergens in latex extracts from 7 different manufacturers and to assess its relevance in the diagnosis of latex allergy. METHODS: Seven latex SPT extracts were analyzed for protein content using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The 4 major allergens Hev b 1, Hev b 3, Hev b 5, and Hev b 6.02 were also quantified using enzyme immunoassay. All commercial extracts were tested for their in vitro allergenic capacity using microarray inhibition assays and for their ability to induce biological reactivity in latex-allergic patients undergoing SPT. RESULTS: The protein content of the extracts varied widely from 8.0 microg/mL to 526.5 microg/mL. SDS-PAGE revealed broad differences in protein profiles between the extracts. Marked variability in the contents of all 4 major allergens was observed, and Hev b 3 and Hev b 5 were undetectable in some extracts. Microarray inhibition assays and SPT demonstrated relevant differences in allergenic capacity between the extracts. CONCLUSIONS: The marked heterogeneity in protein and allergen content of latex extracts from different manufacturers could explain the broad spectrum of SPT results recorded. Our findings suggest that the extracts used for the diagnosis of latex allergy should be improved and standardized.

Allergic hypersensitivity to cannabis in patients with allergy and illicit drug users

Background: Cannabis is the illicit drug most widely used by young people in high-income countries. Allergy symptoms have only occasionally been reported as one of the adverse health effects of cannabis use. Objectives: To study IgE-mediated response to cannabis in drug users, atopic patients, and healthy controls. Methods: Asthmatic patients sensitised to pollen, and all patients sensitised to tobacco, tomato and latex, considered as cross-reacting allergens, were selected from a data base of 21,582 patients. Drug users attending a drug-rehabilitation clinic were also included. Controls were 200 non-atopic blood donors. Specific IgE determination, prick tests and specific challenge with cannabis extracts were performed in patients and controls. Results: Overall, 340 patients, mean age 26.9±10.7 years, were included. Males (61.4%) were the most sensitised to cannabis (p<0.001). All cannabis-sensitised patients were alcohol users. Eighteen (72%) of the patients allergic to tomato were sensitised to cannabis, but a positive specific challenge to cannabis was highest in patients sensitised to tobacco (13/21, 61.9%), (p<0.001). Pollen allergy was not a risk factor for cannabis sensitisation. Prick tests and IgE for cannabis had a good sensitivity (92 and 88.1%, respectively) and specificity (87.1 and 96%) for cannabis sensitisation. Conclusions: Cannabis may be an important allergen in young people. Patients previously sensitised to tobacco or tomato are at risk. Cannabis prick tests and IgE were useful in detecting sensitisation (AU)

Allergic hypersensitivity to cannabis in patients with allergy and illicit drug users.

BACKGROUND: Cannabis is the illicit drug most widely used by young people in high-income countries. Allergy symptoms have only occasionally been reported as one of the adverse health effects of cannabis use. OBJECTIVES: To study IgE-mediated response to cannabis in drug users, atopic patients, and healthy controls. METHODS: Asthmatic patients sensitised to pollen, and all patients sensitised to tobacco, tomato and latex, considered as cross-reacting allergens, were selected from a data base of 21,582 patients. Drug users attending a drug-rehabilitation clinic were also included. Controls were 200 non-atopic blood donors. Specific IgE determination, prick tests and specific challenge with cannabis extracts were performed in patients and controls. RESULTS: Overall, 340 patients, mean age 26.9±10.7 years, were included. Males (61.4%) were the most sensitised to cannabis (p<0.001). All cannabis-sensitised patients were alcohol users. Eighteen (72%) of the patients allergic to tomato were sensitised to cannabis, but a positive specific challenge to cannabis was highest in patients sensitised to tobacco (13/21, 61.9%), (p<0.001). Pollen allergy was not a risk factor for cannabis sensitisation. Prick tests and IgE for cannabis had a good sensitivity (92 and 88.1%, respectively) and specificity (87.1 and 96%) for cannabis sensitisation. CONCLUSIONS: Cannabis may be an important allergen in young people. Patients previously sensitised to tobacco or tomato are at risk. Cannabis prick tests and IgE were useful in detecting sensitisation.

Diagnostic value of Alt a 1, fungal enolase and manganese-dependent superoxide dismutase in the component-resolved diagnosis of allergy to Pleosporaceae.

BACKGROUND: Over the last decades, genomics and proteomics have contributed to the current knowledge of individualized allergenic components and their potential use in the diagnosis of IgE-mediated allergies. Recent investigations have demonstrated that Alt a 1 should be considered as a relevant allergen of the Pleosporaceae group and that enolase is the main allergen involved in the cross-reactivity to fungi. However, the real utility of these allergens as tools for the diagnosis of allergy to Alternaria is still unclear. OBJECTIVE: To demonstrate the current value of the available fungal allergen panel and the need to build an accurate mould allergen array for the diagnosis of allergy to Pleosporaceae. METHODS: Specific IgEs to individual mould allergens and allergenic mould extracts were evaluated using the ImmunoCAP™ system in 30 patients allergic to Alternaria and in 100 blood donors. Cross-reactivity studies were performed by Fluoro Enzyme ImmunoAssay (FEIA) and FEIA inhibition using individual allergens and allergenic extracts. Two-dimensional electrophoresis associated with a MALDI-TOF analysis was carried out to identify new allergen molecules. RESULTS: All allergic patients had positive specific IgE responses to several moulds from different taxonomical families. Classic and molecular diagnosis demonstrated that 23% of patients had multi-sensitization. The current commercially available fungal allergen array was not sufficient to establish an accurate diagnosis. Unexpected correlations between Alternaria or Alt a 1 and Curvularia or Cladosporium stimulated the investigation of a more accurate allergen panel. A manganese-dependent superoxide dismutase (MnSOD) homologous to Asp f 6 was identified as a new IgE-binding molecule from Alternaria alternata. CONCLUSIONS AND CLINICAL RELEVANCE: Alt a 1 is the marker for allergy to Pleosporaceae, not including Curvularia. MnSOD can explain 6.6% of allergy to Alternaria without Alt a 1 sensitization and should be included together with Alt a 1 and fungal enolase in the molecular array for the diagnosis of allergy to Pleosporaceae.

Contribution of molecular diagnosis of allergy to the management of pediatric patients with allergy to pollen.

BACKGROUND: Component-resolved diagnosis based on recombinant allergens facilitates treatment of multiple sensitization and/or crossreactivity in allergic patients. OBJECTIVE: To assess the usefulness of molecular diagnosis in childhood allergies. METHODS: A total of 162 children aged 4-16 years diagnosed with allergic rhinitis or asthma/rhinitis caused by pollen were referred for recombinant allergen-based diagnosis in 2006. Specific immunoglobulin (Ig) E against pollen allergens and purified recombinant Phleum pratense pollen allergens were measured using an in vitro quantitative assay, and considering the recombinant allergens Phl p 1+Phl p 5 as P pratense--specific allergens and Phl p 7+Phl p 12 as cross-reacting allergens. Conditional probability was calculated to determine the relationship between values for specific IgE against major allergens and those for cross-reacting allergens. RESULTS: Specific IgE antibodies against P pratense were detected in 99.4% of serum samples, and cross-reacting allergens in 46%. Multiple sensitization to pollen was documented in 38% of patients, with Plantago lanceolata as the main cause. Conditional probability calculations showed that patients with specific IgE values of 75-80 kU(A)/L to Phl p 1+Phl p 5 were 75% (95% confidence interval) more likely to present values > or = 2 kUA/L to Phl p 7+Phl p 12. CONCLUSIONS: Our results show that recombinant DNA technology can help diagnose allergy in cases of multiple sensitization and crossreactivity, and is therefore a promising option for improving prognosis and management of allergic pediatric populations.

Identification of 2 new allergens of Phoenix dactylifera using an immunoproteomics approach.

The date palm (Phoenix dactylifera) has a wide geographical distribution (Middle East, Mediterranean, central Africa, western Asia, Australia, and North America). Pho d 2, the major allergen of date palm pollen was recently identified as a profilin, yet little is known about the nature of the other pollen allergens from this tree. The objective of this study was to characterize clinically significant allergens other than profilins from P. dactylifera pollen using immunoproteomics. In order to reveal the proteins causing the allergy, we used serum from a patient monosensitized to date palm pollen extract who experienced asthma and rhinoconjunctivitis during the palm tree pollen season. The results revealed 2 novel immunoglobulin E-binding proteins not related to the cross-reactive allergen profilin. Individualized allergens of Pdactylifera that cause specific date palm pollen sensitization must be defined to determine the real prevalence of sensitization to this species.

Síndrome ave-huevo / Bird-egg syndrome

Antecedentes. Se ha descripto una relación entre la hipersensibilidad respiratoria tipo I frente a antígenos aviares y la alergia alimentaria a la yema de huevo. Dicha asociación se denomina síndrome ave-huevo, y el responsable de dicho cuadro es la alfa-livetina o seroalbúmina de pollo, un antígeno presente tanto en la yema del huevo como en las plumas, suero y excrementos de las aves. Materiales y métodos. Estudiamos una paciente con síntomas de alergia alimentaria tras la ingesta de huevo, quien además sufría de síntomas respiratorios (rinitis/asma) causados por la exposición a aves. Se realizaron pruebas cutáneas con huevo, alfa-livetina, pollo crudo y cocido, y plumas. La IgE sérica específica fue identificada por técnica de microarrays de alérgenos (Immuno CAP ISAC). Resultados. Los prick test fueron positivos para alfa-livetina (8 mm), pollo crudo (8 mm) y plumas de gallina (7 mm). La determinación de IgE sérica específica fue de 16,61 (kU/l) para alfa­livetina. Conclusiones. El síndrome ave-huevo es producido por la sensibilización a la alfa livetina, un alérgeno que puede actuar tanto por vía alimentaria como por vía inhalatoria. Según nuestro conocimiento, es el primer caso diagnosticado a través de la técnica de microarray de alérgenos.(AU)

Background: A relationship between type I hypersensitivity with respiratory symptoms due to bird antigens and allergy to egg yolk has been described. This association is known as bird-egg syndrome, which is caused by sensitization to chicken serum albumin (alpha-livetin), present in bird feathers and serum, and egg yolk. Material and methods: We studied one patient with food allergy to egg yolk who also suffered from respiratory symptoms (rhinitis- asthma) caused by exposure to birds. Sensitization to egg yolk and bird antigens was investigated by skin prick test. Specific IgE was investigated using allergens Microarrays (Immuno CAP ISAC). Results:Our patient had a positive skin prick test to: chicken serum albumin (alpha livetin): 8 mm, bird feathers: 7 mm, raw chicken: 8 mm. Specific IgE to alpha livetin was 16.61 (kU/l). Conclusions: Bird-egg syndrome is due to a sensitization to alpha-livetin, an allergen that can act either on the respiratory or the digestive way. In our knowledgement, this is the first case described using allergen Microarrays technique.(AU)

Galectin and aldolase-like molecules are responsible for the specific IgE response in humans exposed to Dirofilaria immitis.

Dirofilaria immitis is the agent of the heartworm disease in canids and felids, and of pulmonary dirofilariosis in man. Like other filariae, D. immitis harbours endosymbion Wolbachia bacteriae. In this work we analyse the response of specific IgE antibodies against both D. immitis antigens and the Wolbachia surface protein (WSP) in two groups of persons living in an area of canine endemia, one presenting high levels of total IgE (group 1) and other with normal levels (group 2). Infections with D. immitis were demonstrated by the presence of specific IgG in 228 individuals(48.8%) of the group 1 and only in one of the group 2. Specific IgE antibody response against D. immitis antigens was detected only in individuals of the group 1. IgE response against WSP was not detected in any group. The IgE response was directed mainly against two molecules of 33 and 42 kDa of the antigenic extract of D. immitis. These molecules were identified by mass spectrometry as a galectin and an aldolase, respectively. Their possible role in the survival mechanisms of the parasite and their contribution to development of allergic reactions in individuals resident in areas with heartworm disease are discussed.

Variability of Alt a 1 expression by different strains of Alternaria alternata.

BACKGROUND: While it is well known that there is significant intraspecific variation in the content an potency of Alternaria alternata allergens, little data has been published on intraspecific variability for individual allergens from moulds. OBJECTIVE: To assess the variability of Alt a 1 expression in different strains of A alternata. METHODS: Eleven strains of A alternata were cultured in a Czapek broth medium and culture filtrate extracts were obtained. A sensitive two-site enzyme-linked immunosorbent assay was used to measure Alt a 1 concentrations in medium following 3 weeks of culture and in culture filtrate extracts. RESULTS: Expression of Alt a 1 was highly variable in different strains ofA alternata (coefficient of variation > 135%). A good correlation was found between Alt a 1 concentrations at the beginning of the process and measurements at the end of extract production (r=0.940). CONCLUSIONS: The high variability of Alt a 1 expression in different A alternata strains makes it necessary to measure Alt a 1 concentrations during the first stage of allergenic extract production in order to be able to choose a suitable strain for producing extracts or purifying Alt a 1.

Uncinaria stenocephala: assessment of antigens for the immunodiagnosis of canine uncinariosis.

Although Uncinaria stenocephala is the most frequent hookworm in the intestine of dogs from Northern, Central and Southern Europe, little is known about its host-parasite relationship. Three groups of sera from dogs (Group 1: dogs naturally infected only by U. stenocephala; Group 2: helminth-free dogs at necropsy, and Group 3: dogs parasitized by other helminths) were analyzed by ELISA using U. stenocephala antigens from adult worms (somatic and excretory-secretory antigens) and from L3 larvae (somatic antigens). All three sources of antigens were found to be suitable for immunodiagnosis of canine uncinariosis with up to 90% efficacy. However, an analysis to assess the diagnostic value of the different antigens demonstrated that the adult excretory-secretory antigens had a higher diagnostic efficacy (96.7%), indicating that this is the best antigen source for the diagnosis of Uncinaria infection.

Barnacle allergy: allergen characterization and cross-reactivity with mites.

BACKGROUND: Barnacles are a type of seafood with worldwide distribution and abundant along the shores of temperate seas. They are particularly appreciated and regularly consumed in Portugal as well as in Spain, France and South America, but barnacle allergy is a rare condition of which there is only one reference in the indexed literature. The molecular allergens and possible cross-reactivity phenomena implicated (namely with mites) have not been established. OBJECTIVE: To demonstrate the IgE-mediated allergy to barnacle and to identify the proteins implicated as well as possible cross-reactivity phenomena with mites. METHODS: We report the clinical and laboratory data of five patients with documented IgE-mediated allergy to barnacle. The diagnosis was based on a suggestive clinical history combined with positive skin prick tests (SPT) to barnacle--prick to prick method. Two barnacle extracts were prepared (raw and cooked barnacle) and sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) and IgE-immunoblotting were performed. An immunoblotting inhibition assay with Dermatophagoides pteronyssinus was also done in order to evaluate cross-reactivity. RESULTS: All patients had mite-related asthma and the allergic rhinoconjunctivitis; they all experienced mucocutaneous symptoms. All of them had positive SPT to barnacle, and the immunoblotting showed several allergenic fractions with a wide molecular weight range (19 - 94 kDa). The D. pteronyssinus extract inhibited several IgE-binding protein fractions in the barnacle extract. CONCLUSIONS: We describe five patients with IgE-mediated barnacle allergy. We also describe a group of IgE-binding+proteins between 30 and 75 kDa as the allergenic fractions of this type of Crustacea. Cross-reactivity with D. pteronyssinus was demonstrated in two cases.