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PubMed2002

Specific IgE determination to Ani s 1, a major allergen from Anisakis simplex, is a useful tool for diagnosis.

BACKGROUND: The most serious limitation of the serodiagnosis of parasitoses is the occurrence of cross-reactions. OBJECTIVE: The possible use of Anisakis simplex major allergen Ani s 1 for diagnosis. PATIENTS AND METHODS: Forty-nine non-fish-allergic patients with Anisakis simplex hypersensitivity, 21 patients without allergic episodes suffering intestinal anisakiasis with obstruction of the intestinal lumen, and 10 unrelated sera as a control were included in this study to determine specific immunoglobulin (Ig)E and IgG to Anisakis simplex major allergen Ani s 1 by immunoblotting. RESULTS: Eighty-six percent of patients with Anisakis simplex hypersensitivity showed specific IgE directed to Ani s 1. Identical result was obtained for IgG detection in this group. Among patients with intestinal anisakiasis, 86% showed specific IgE, but only 29% had specific IgG (P < 0.001, two-tailed Fisher exact test). One of the 10 control subjects was positive both for IgE and IgG (P < 0.001). CONCLUSIONS: Determination of specific IgE directed to Anisakis simplex major allergen Ani s 1 is a useful tool for the diagnosis of hypersensitivity and intestinal anisakiasis. Further, measurement of specific IgG directed to Anisakis simplex major allergen Ani s 1 is only valid for Anisakis simplex allergy.

Allergens↗
PubMed2000

Incidence of sensitivity to Anisakis simplex in a risk population of fishermen/fishmongers.

BACKGROUND: Anisakis simplex, a fish and cephalopodes parasite, can cause either gastrointestinal symptoms or allergic reactions in humans on eating/handling contaminated fish. OBJECTIVE: The aim of our study was to determine the capacity of Anisakis simplex to induce specific IgE production and allergic reactions following eating and handling fish in a population at risk. METHODS: We determined the levels of total IgE, specific IgE, and eosinophil count in 28 fishermen/fishmongers (group A) and 15 healthy donors (group B). A skin prick test (SPT) with extracts from Anisakis and the most common species of fish in our country, were also carried out. RESULTS: Specific IgE to Anisakis were found in 14 subjects of group A (13 of them had a positive SPT to the same extract) and none of group B (only one subject had a positive SPT). The SPT with fish extracts was positive in 4 patients of group A but in none of group B. Subjects in group A with specific IgE to Anisakis showed higher total IgE levels and eosinophil counts compared with either other individuals of the same group or to those of group B. CONCLUSIONS: These results indicate that fishermen/fishmongers are a population at risk for Anisakis simplex sensitization and suggest that this kind of sensitization should also be investigated not only in subjects like fishermen/fishmongers who live in countries where fish is likely to be contaminated with Anisakis simplex parasites, but also in those who handle fish for other reasons.

Adolescent↗
PubMed2006

Genetic relationships among Anisakis species (Nematoda: Anisakidae) inferred from mitochondrial cox2 sequences, and comparison with allozyme data.

The genetic relationships among 9 taxa of Anisakis Dujardin, 1845 (A. simplex (sensu stricto), A. pegreffii, A. simplex C., A. typica, A. ziphidarum, A. physeteris, A. brevispiculata, A. paggiae, and Anisakis sp.) were inferred from sequence analysis (629 bp) of the mitochondrial cox2 gene. Genetic divergence among the considered taxa, estimated by p-distance, ranged from p = 0.055, between sibling species of the A. simplex complex, to p = 0.12, between morphologically differentiated species, i.e., A. ziphidarum and A. typica. The highest level was detected when comparing A. physeteris, A. brevispiculata, and A. paggiae versus A. simplex complex (on average p = 0.13) or versus A. typica (on average p = 0.14). Sequence data from the newly identified Anisakis sp. poorly aligned with other Anisakis species but was most similar to A. ziphidarum (p = 0.08). Phylogenetic analyses based upon Parsimony and Bayesian Inference, as well as phenetic analysis based upon Neighbor-Joining p-distance values, generated similar tree topologies, each well supported at major nodes. All analyses delineated two main claides, the first encompassing A. physeteris, A. brevispiculata, and A. paggiae as a sister group to all the remaining species, and the second comprising the species of the A. simplex complex (A. simplex (s.s.), A. pegreffii and A. simplex C), A. typica, A. ziphidarum, and Anisakis sp. In general, mtDNA-based tree topologies showed high congruence with those generated from nuclear data sets (19 enzyme-loci) and with morphological data delineating adult and larval stages of the Anisakis spp.; however, precise positioning of A. typica and A. ziphidarum remain poorly resolved, though they consistently clustered in the same clade as Anisakis sp. and the A. simplex complex. Comparison of anisakid data with those currently available for their cetacean-definitive hosts suggests parallelism between host and parasite phylogenetic tree topologies.

Animals↗
PubMed2006

[A case of immediate allergy to anisakis following saury intake].

We report here a 76-year-old male that presented with an immediate allergy to Anisakis following saury intake. Three and a half hours after eating pressed saury sushi, whole-body pomphus appeared including itching, facial dropsical swelling, and dyspnea. Diagnostic tests revealed specific IgE antibodies against anisakis simplex and a skin prick test was positive using an extraction of anisakis simplex. The results of skin prick tests using the body and internal organs of a saury were negative. Based on these results, we diagnosed the case as immediate allergy to Anisakis. Anisakis is parasitic to a diverse array of fish, and it seems rare that eating saury will induce an allergic response because the reported parasitic rate of Anisakis on saury is only 5%. In addition, as tropomyosin is currently considered to be the primary cause of allergies to Anisakis, renewed attention should be paid to other foods for which tropomyosin is also assumed to be a common antigen.

Aged↗
PubMed2000

Genetic markers in ribosomal DNA for the identification of members of the genus Anisakis (Nematoda: ascaridoidea) defined by polymerase-chain-reaction-based restriction fragment length polymorphism.

Polymerase-chain-reaction-based restriction fragment length polymorphism analysis was performed to establish genetic markers in rDNA, for the identification of the three sibling species of the Anisakis simplex complex and morphologically differentiated Anisakis species, i.e. Anisakis physeteris, Anisakis schupakovi, Anisakis typica and Anisakis ziphidarum. Different restriction patterns were found between A. simplex sensu stricto and Anisakis pegreffii with two of the restriction endonucleases used (HinfI and TaqI), between A. simplex sensu stricto and A. simplex C with one endonuclease (HhaI), and between A. simplex C and Aniskis pegreffii with three endonucleases (HhaI, HinfI and TaqI), while no variation in patterns was detected among individuals within each species. The species A. physeteris, A. schupakovi, A. typica and A. ziphidarum were found to be different from each other and different from the three sibling species of the A. simplex complex by distinct fragments using 10-12 of the endonucleases tested. The polymorphisms obtained by restriction fragment length polymorphisms have provided a new set of genetic markers for the accurate identification of sibling species and morphospecies.

Animals↗
PubMed1990

[Analysis of antigens defined by anti-Anisakis larvae antibodies of IgE and IgG type in the sera of patients with acute gastrointestinal anisakiasis].

Using SDS-PAGE and Immunoblotting assay, we analysed antigens defined by anti-Anisakis Larvae antibodies of IgE and IgG type in the sera of patients with acute gastrointestinal anisakiasis. 1) Humoral IgE response to 76, 64 and 60 kDa molecular weight proteins of Anisakis Larvae and IgG response to 91, 76, 64 and 60 kDa molecular weight proteins were demonstrated in the sera of patients with acute gastric anisakiasis. Those humoral antibody responses were also demonstrated in the sera of two patients with intestinal anisakiasis. 2) Those antibodies to 76, 64 and 60 kDa proteins may be Anisakis-specific antibodies, because anti-Anisakis Larvae antibodies of IgE and IgG type did not react with the similar MW proteins of Ascaris suum homogenates. 3) Also anti-Anisakis Larvae antibodies reacted with Anisakis Excretory and Secretory (E.S.) protein which included specific 76 and 60 kDa proteins, but they did not react with purified Anisakis hemoglobin.

Acute Disease↗
PubMed1999

Human immunoglobulin isotype profiles produced in response to antigens recognized by monoclonal antibodies specific to Anisakis simplex.

BACKGROUND: Anisakis simplex is a medically important pathogen which not only causes anisakiasis but may provoke allergy reactions, ranging from mild urticaria to anaphylactic shock. OBJECTIVE: To investigate anti-Anisakis isotype profiles in anisakiasis and Anisakis allergy patients. METHODS: Capture ELISA techniques were used to investigate the isotype profiles of antibodies specific for two defined Anisakis simplex antigens, in serum from Japanese patients with confirmed anisakiasis and from Spanish patients with allergy to Anisakis. The antigens were 'UA2R antigens' (two proteins with MW of 48 and 67 kDa, recognized by our monoclonal antibody UA2) and 'UA3R antigens' (two proteins with MW of 139 and 154 kDa, recognized by our monoclonal antibody UA3). RESULTS: Considering IgG, the two most frequent isotypes in the response to the UA2R antigens were IgG1 and IgG2, with IgG4 detected in only one case; in response to the UA3R antigens, by contrast, the two most frequent isotypes were IgG1 and IgG4 (though IgG2 remained reasonably frequent). As regards potential utility for serodiagnosis, 95% of the Japanese anisakiasis patients and 84% of the allergy patients showed detectable IgG1 antibodies to the UA3R antigens. Furthermore, all allergy patients showed IgE antibodies to these antigens. CONCLUSION: Anisakis simplex contains antigens that induce responses which are differentially regulated. Because of their immunogenicity, immunodominance and allergenic nature, we consider that the 139/154-kDa antigens recognized by our MoAb UA3 are good candidates for use in tests for the diagnosis of anisakiasis and of the allergy caused by this parasite.

Anaphylaxis↗
PubMed2006

Molecular systematics, phylogeny and ecology of anisakid nematodes of the genus Anisakis Dujardin, 1845: an update.

Advances in the taxonomy and ecological aspects concerning geographical distribution and hosts of the so far genetically recognised nine taxa of the nematodes belonging to genus Anisakis (i.e. A. pegreffii, A. simplex s.s., A. simplex C, A. typica, A. ziphidarum, Anisakis sp., A. physeteris, A. brevispiculata and A. paggiae) are here summarized. Genetic differentiation and phylogenetic relationships inferred from allozyme (20 enzyme-loci) and mitochondrial (sequences of cox-2 gene) markers, are revised and compared. The two genetic analyses are congruent in depicting their phylogenetic relationships. Two main clusters are showed to exist in the obtained trees, one encompassing the species A. pegreffii, A. simplex s.s., A. simplex C, A. typica, A. ziphidarum and Anisakis sp.; while, the second including A. physeteris, A. brevispiculata and A. paggiae. The existence of two clades is also supported by their morphological differentiation in adult and larval morphology. Comparison of phylogenetic relationships among Anisakis spp. with those currently available for their cetacean definitive hosts suggests parallelism between host and parasite phylogenetic tree topologies. Preliminary data for reconstruction of a possible co-evolutionary scenario between cetacean hosts and their Anisakis endoparasites suggests that cospeciation and host-switching events may have accompanied the evolution of this group of parasites. Finally, genetic/molecular markers for the identification of the so far genetically recognized taxa of Anisakis at any life-stage and both sexes were given also in relation to human anisakiosis is discussed.

Animals↗
PubMed2006

Molecular identification of Anisakis species from Pleuronectiformes off the Portuguese coast.

Anisakid nematodes belonging to the Anisakis simplex complex are highly prevalent in several fish species off the coast of Portugal and are an important zoonotic problem in the Iberian Peninsula. Two reproductively isolated sibling species of the Anisakis simplex complex were identified from Pleuronectiformes inhabiting the Portuguese coast using restriction fragment length polymorphism (RFLP). Recombinant genotypes corresponding to presumptive Anisakis simplex sensu stricto and Anisakis pegreffii hybrids were also detected by this technique, as well as the species Anisakis typica. Although 25 species of Pleuronectiformes were investigated, Anisakis spp. larvae were only found in seven: Arnoglossus imperialis, Arnoglossus laterna, Lepidorhombus boscii, Citharus linguatula, Platichthys flesus, Dicologlossa cuneata and Solea senegalensis. The occurrence of hybrids in relatively sedentary fishes such as the Pleuronectiformes suggests that the Portuguese coast may constitute an area of hybridization and, therefore, is of particular interest for the study of the process of hybridization and speciation for these anisakids.

Animals↗
PubMed2005

Detection of Anisakis simplex-induced basophil activation by flow cytometry.

BACKGROUND: Laboratory diagnosis of anisakidosis is based on specific serum IgE detection. Recently, detection of allergen-induced basophil activation by flow cytometry has been proposed as a valuable tool for allergy diagnosis. OBJECTIVE: To evaluate if detection of Anisakis-induced basophil activation by flow cytometry is a useful tool in the diagnosis of Anisakis allergy. METHODS: Patients with Anisakis allergy (A.s.+, n = 37), patients reporting chronic urticaria or abdominal pain unrelated to fish ingestion (A.s.-, n = 51), and healthy controls (n = 12) were studied. Specific IgE to Anisakis simplex (A. simplex) was quantified with CAP-FEIA method, and basophil activation test was performed with three different concentrations of an Anisakis crude extract. Basophil gating was performed with CD123 and HLA-DR, and cellular activation was measured with CD63. RESULTS: A.s.+ patients showed significantly higher age and total IgE levels than did the A.s.- patients. Specific IgE to A. simplex correlated with the activated basophil percentages obtained with 15 microg/mL (r = 0.80; P < 0.001), 1.5 microg/mL (r = 0.84; P < 0.001), and 0.15 microg/mL (r = 0.82; P < 0.001) of A. simplex crude extract. Nine individuals (3 in the A.s.+ group and 6 in the A.s.- group) were nonresponders to basophil stimulation with anti-IgE. Five A.s.- patients showed positive IgE values to A. simplex while the basophil activation test was negative. According to the receiver operating characteristics curves performed between A.s.+ vs. A.s.- and A.s.+ vs. healthy controls, the cutoff for a positive basophil activation test was >or=21% (specificity = 96%, sensitivity = 100%), and 16% (sensitivity and specificity of 100%) respectively. When nonresponders were included in the A.s.+ vs. A.s.- analysis, sensitivity decreased to 95%. Multivariate logistic analysis showed that the specific basophil activation was a factor independently associated with clinical symptoms of A. simplex allergy. CONCLUSIONS: Detection of A. simplex-induced basophil activation by flow cytometry is a useful laboratory technique for the diagnosis of anisakidosis, supplementing specific IgE determinations.

Adult↗
PubMed1997

Allergic reactions to anisakis simplex parasitizing seafood.

BACKGROUND: The ingestion of Anisakidae ssp larvae parasitized fish can cause anisakiasis. Allergic reactions after ingestion of safely cooked but parasitized fish have been reported. METHODS: Twenty-three patients who suffered allergic reactions after seafood ingestion, with negative skin tests were studied. Anisakis simplex sensitization was assessed by skin prick test and/or specific serum Immunoglobulin E (IgE). Total serum IgE and specific IgE against the implicated seafood and Ascaris lumbricoides were also determined. RESULTS: Manifestations of Anisakis simplex allergy were urticaria/angioedema (18/23) patients and anaphylaxis (5/23). Gastric symptoms were also observed (3/23). Sea fish and shellfish were implicated. Raw and cooked seafood ingestion caused reactions. Total serum IgE ranged from 13 to 7200 KU/L. Specific IgE to Anisakis simplex was positive (> 0.35 KU/L) in all patients, and skin tests were positive in 20. Serum-specific IgE and skin tests to the involved seafoods were negative in every patient. Serum-specific IgE to Ascaris lumbricoides was negative in 13 patients. No association between total IgE and the eosinophil count (r < 0.1) was observed, but there was some association between total IgE and specific IgE to Anisakis simplex (r = 0.58). CONCLUSION: Anisakis simplex sensitization is the cause of allergic reactions after seafood ingestion. It is important to pay attention to this new "food allergy" to diagnose correctly the etiology of adverse food reactions.

Adolescent↗
PubMed2002

Acute allergic reactions to Anisakis simplex after ingestion of anchovies.

Anisakis simplex is a seafish nematode, which is responsible for the well-known human infection (anisakiasis) and can induce IgE-mediated reactions. IgE sensitization to Anisakis simplex can be frequent in particular countries and should be suspected in patients with acute allergic symptoms after ingestion of fish. The etiological role of Anisakis simplex was evaluated in 49 adult subjects with acute allergic symptoms after ingestion of anchovies. Serum-specific IgE and prick tests to anchovy were negative in each patient. Specific IgE reactions to Anisakis were positive in 45 patients and skin tests in 43. Only 3 patients with allergy to the nematode were atopic. However, IgE responses to Anisakis were also observed in habitual consumers of raw fish, without any clinical manifestations, suggesting that the relevance of results of conventional tests has to be interpreted on the basis of clinical aspects.

Acute Disease↗
PubMed1997

Cross-reactivity between IgE-binding proteins from Anisakis, German cockroach, and chironomids.

Anisakis simplex larvae parasitize animals used as seafood and can produce a specific immune response in man. The ingestion of seafood contaminated with stage three of A. simplex larvae can induce a specific IgE response with clinical symptoms, usually urticaria, even if the fish is cooked before ingestion and the invasive infestation power destroyed by heating. Our preliminary studies showed a strong association of A. simplex sensitization with Ascaris lumbricoides, Daphnia, chironomid spp., Atlantic shrimp (Pandalus borealis), and German cockroach (Blattella germanica). We conducted the cross-reactivity study with cockroach, a ubiquitous insect, and Chironomidae (red mosquito larvae), a work-related allergen, without any possibility of Anisakis contamination. Serum samples were collected from 60 pediatric patients, with serum specific IgE to A. simplex. Both specific-IgE and immunoblot-inhibition studies, with a serum pool from 18 patients, were performed to determine whether the association of sensitizations to nematodes and arthropods was due to immunologic cross-reactivity. In addition, serum samples from 21 of 60 patients who showed also sensitization to German cockroach were used for individual immunoblot studies. In the serum pool, dose-dependent inhibition of B. germanica and Chironomus spp, was observed after preincubation with the A. simplex extract. Immunoblot of Anisakis, inhibited with Chironomus and German cockroach, yielded a partial blot inhibition but mainly on bands below 41 kDa. Blot inhibition of German cockroach and Chironomus with Anisakis was dose related. The band patterns in individual blots were heterogeneous, but most of them had bands of 30-43 kDa. None of these sera recognized allergens in the 14-kDa area. In our study, CAP-inhibition and immunoblot-inhibition analysis of Anisakis showed that several IgE-binding components could be shared by the three allergens.

Allergens↗
PubMedDate not supplied

Risk factors for sensitization to AnisakiS simplex: a multivariate statistical evaluation.

Anisakis simplex is a nematode belonging to the Anisakidae family. The ingestion of third stage larvae in uncooked or undercooked seafood may cause human diseases known as anisakiasis and anisakidosis. A total of 400 (159 atopic and 241 non-atopic) subjects living in an area of southern Italy (Bari district) were consecutively evaluated to identify the association of some factors (sex, age, atopy, consumption of uncooked seafood and sensitization to dust mites) with the risk of Anisakis simplex sensitization. Patients were investigated on history of atopy and allergic diseases and were skin prick tested with commercial allergen extracts of Anisakis simplex, Acarus siro, Lepidoglyphus destructor, Tyrophagus putrescentiae, Glycyphagus domesticus, Euroglyphus maynei, Dermatophagoides pteronyssinus and Dermatophagoides farinae. Our results suggest that atopic subjects have a lower risk of Anisakis allergy than non-atopic subjects and show the association of Anisakis simplex sensitization with the consumption of uncooked seafood (anchovies and squid), increasing age and sensitization to Glycyphagus domesticus.

Animals↗
PubMed2000

Anisakis simplex: a cause of intestinal pseudo-obstruction.

OBJECTIVE: the ingestion of Anisakis simplex larvae may lead to the appearance of gastrointestinal symptoms. However, the number of reported cases of parasitization by Anisakis in Spain is lower than would be expected in a country with the second-highest fish consumption per inhabitant in the world, particularly since fish is often eaten raw or only slightly cooked. We suggest that the incidence of anisakiasis in Spain would be higher if complementary studies were used in all patients suspected of having anisakiasis. METHODS: we studied 6 patients with a diagnosis of intestinal obstruction who frequently ate fish. Skin prick tests with seafood, inhalant allergen and Anisakis extracts were done. Total and specific IgE against Anisakis larvae were tested with a CAP system radioimmunoassay and immunoblot assays. Oral challenge tests with frozen larvae were also used. RESULTS: a positive skin prick result and high levels of total and specific IgE were found in all patients. The results of immunoblot assays for IgE did not show a consistent pattern, but a group of several low (14-18 kDa) and intermediate molecular weight antigens (30-50 kDa) were found in all patients. All patients tolerated the oral challenge test well. CONCLUSIONS: in our patients with intestinal pseudo-obstruction and a history of frequent fish eating, the clinical and laboratory findings were suggestive of parasitization by Anisakis simplex larvae as the cause of the obstruction. Such complementary studies should be used whenever there is a suspicion of anisakiasis. The results of the oral provocation test show that the intake of dead larvae does not induce clinical parasitization.

Abdomen, Acute↗
PubMed2000

[Eosinophilic esophagitis associated with recurrent urticaria: is the worm Anisakis simplex involved?].

Anisakis simplex, a fish parasite of the nematode family, typically infects marine mammals such as whales, dolphins and seals. Human anisakiasis, which is acquired by eating raw or insufficiently heated fish or squid, has gained world-wide importance. Infestation with living larvae caused by eating parasitised fish results in acute upper abdominal pain, nausea and vomiting and may be confused with acute abdomen due to appendicitis and other inflammatory abdominal disorders. Extraintestinal organ manifestations are rare. Endoscopically, inflammation, oedema, erosions and ulcerations may be found. The parasite can been found in up to 50% of patients. Histologically, an eosinophilic inflammation is typical. Acute anisakiasis may be prevented by thorough cooking or deep-freezing the parasitised fish for at least 48 h. IgG-antibodies specific for Anisakis simplex are thought to represent an immunological host reaction against parasitic antigens. More recently, allergic reactions to Anisakis ingestion or exposure, such as urticaria, anaphylaxis and even occupational asthma, have been reported. These allergic reactions may also occur when the fish has been properly cooked, and hence these allergens are thought to be heat-stable. Such cases may be diagnosed by skin tests and the determination of specific Anisakis-IgE. However, the specificity of IgE is low, since they may also be present in exposed asymptomatic individuals. Since the eliciting allergens are temperature-stable, prophylactic dietetic measures are indicated. We report a case from Switzerland acquired during a holiday in Portugal. The patient suffered from recurrent dysphagia and urticaria, and histologically eosinophilic oesophagitis was found. IgG-antibodies and a positive skin prick test to Anisakis simplex support its aetiologic role for the symptoms.

Adult↗
PubMed2006

Molecular cloning and expression of two new allergens from Anisakis simplex.

The nematode Anisakis simplex is a marine parasite that causes allergy as well as anisakiasis. Although five Anisakis allergens have already been identified, immunoblotting studies suggested that unidentified allergens still exist. In this study, an expression cDNA library constructed from A. simplex was subjected to immunoscreening using an Anisakis-allergic patient serum, and two positive clones coding for allergens (named Ani s 5 and 6) were obtained. Ani s 5 (152 amino acid residues) is homologous with nematode proteins belonging to the SXP/RAL-2 protein family and Ani s 6 (84 amino acid residues) with serine protease inhibitors from various animals. Of the 28 patient sera examined, seven and five reacted to recombinant Ani s 5 and 6 expressed in Escherichia coli, respectively. By inhibition immunoblotting experiments using the recombinant allergens as inhibitors, natural Ani s 5 could be identified as a 15-kDa protein in the crude extract of A. simplex but natural Ani s 6 could not be identified probably due to its low expression. In conclusion, Ani s 5 and 6 are new allergens of A. simplex that are specific to some Anisakis-allergic patients.

Allergens↗
PubMed2001

Purification of Anisakis simplex antigen by affinity chromatography.

In order to improve the specificity and sensitivity of the techniques for the diagnosis of human anisakidosis, a method of affinity chromatography for the purification of species-specific antigens from Anisakis simplex third-stage larvae (L3) has been developed. New Zealand rabbits were immunized with A. simplex or Ascaris suum antigens or inoculated with Toxocara canis embryonated eggs. The IgG-specific antibodies were isolated by means of protein A-Sepharose CL-4B bead columns. IgG anti-Anisakis simplex, anti-Ascaris suum and anti-T. canis were coupled to CNBr-activated Sepharose 4B. For the purification of the larval Anisakis simplex antigens, it was loaded into the anti-A. simplex column and bound antigens were eluted. For the elimination of the epitopes responsible for the cross-reactions, the A. simplex-specific proteins were loaded into the anti-Ascaris suum and anti- T. canis columns. To prove the specificity of the isolated proteins, immunochemical analyses by polyacrylamide gel electrophoresis and immunoblotting were carried out. Likewise, immunoaffinity columns were prepared using specific IgG from patients with Anisakis simplex sensitization, previously diagnosed by fluoro-enzymo-immunoassay. The protein patterns of antigen after purification by the human columns were similar to those obtained using the rabbit columns.

Animals↗