Enzyme-linked immunosorbent assay and Western blot antibody determination in sera from patients diagnosed with different helminthic infections with Anisakis simplex antigen purified by affinity chromatography

Rodero, M.; Chivato, T.; Muro, Antonio; Cuéllar, C.

doi:10.1590/s0074-02762005000300015

Cited by 6 publications

(9 citation statements)

References 26 publications

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“…The results of this fluoroimmunoassay were reported in ku/ℓ and converted to CAP ‘scores’ of 0( < 0.35), 1(0.35–0.7), 2(0.7–3.5), 3(3.5–17.5), 4(17.5–50), 5(50–100) and 6(>100). All sera showed positive CAP values to Anisakis by FEIA, which varied from CAP = 2 (41.6%), CAP = 3 (25%), CAP = 4 (12.5%), CAP = 5 (12.5%) to CAP = 6 (8.3%) (Rodero et al , 2005). A column was prepared with Protein A affinity isolated IgG anti- A .…”

Section: Methodsmentioning

confidence: 99%

Western blot antibody determination in sera from patients diagnosed with Anisakis sensitization with different antigenic fractions of Anisakis simplex purified by affinity chromatography

Rodero

Cuéllar

Chivato³

et al. 2007

J. Helminthol.

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Using Western blot techniques, the specificities of crude and purified (PAK and PAS) Anisakis simplex antigens were compared against 24 sera from patients diagnosed with Anisakis sensitization. All patients recognized a 60 kDa protein against the A. simplex crude extract, while 37.5% and 12.5% reacted with proteins of 40 and 25 kDa, respectively, when IgG was tested. In the case of IgE determination, 41.6% of sera were negative, while 12.5% and 20.8% appeared to cross-react against Toxocara canis and Ascaris suum, respectively. When the PAK antigen (A. simplex antigen purified by means of a column of IgG anti-A. simplex) was tested, immune recognition towards the 60, 40 and 25 kDa proteins increased in 83.3%, 16.7% and 4.2%, respectively, when the Ig antibodies were tested. In the case of the PAS antigen (PAK antigen purified by means of a column of IgG anti-A. suum), the reaction against the 40 and 25 kDa proteins increased to 45.8% and 25%, respectively, when Ig antibodies were used. Finally, when the EAS antigen (eluted from the anti-A. suum column after PAK purification) was tested, 83.3% of the assayed sera reacted against the 14 kDa protein, when the Ig antibodies, IgG and IgM immunoglobulins were measured. With the IgE determination, the reactions were observed in 41.7% of patients with proteins between 60 and 35 kDa against the PAS antigen. With the EAS antigen, reactive bands of 184, 84 and 14 kDa appeared. In conclusion, in the purification process of the A. simplex larval crude extract, the proteins implicated in cross-reactions with Ascaris and Toxocara were eliminated, with an important concentration of proteins responsible for the induction of specific responses.

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Section: Methodsmentioning

confidence: 99%

Western blot antibody determination in sera from patients diagnosed with Anisakis sensitization with different antigenic fractions of Anisakis simplex purified by affinity chromatography

Rodero

Cuéllar

Chivato³

et al. 2007

J. Helminthol.

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“…Numerous teleosts also play a part in this life cycle and can carry the infective L3 larvae in the body cavity or muscle, thereby posing a health risk to human consumers of the fish (Klimpel et al , 2008). Anisakis larvae can induce lesions in the human digestive tract and can also produce allergic reactions through the consumption of parasitized fish and, to a lesser degree, through contact or inhalation or cross-reactivity with allergens of other organisms (Daschner et al , 2000; Scala et al , 2001; Rodero et al , 2005; Solas et al , 2009). In addition, from an economic standpoint, Anisakis infection can have a negative impact on fish sales because the larvae are visible to the naked eye.…”

Section: Introductionmentioning

confidence: 99%

Epidemiology and molecular identification ofAnisakis pegreffii(Nematoda: Anisakidae) in the horse mackerelTrachurus trachurusfrom northern Morocco

et al. 2013

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Anisakis infection parameters were studied in horse mackerel (Trachurus trachurus) taken from two areas of northern Morocco (Tetouan and Tangier), which showed a mean prevalence of 54.9%. Identification of the ITS1-5.8S-ITS2 fragment by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) showed A. pegreffii to be the dominant species; no A. simplex s.s. were detected. The presence of A. pegreffii in horse mackerel was not influenced by the sex (P = 0.46) or catch area (Atlantic versus Mediterranean, P = 0.52) of the fish, but was significantly related to their length, weight, liver weight and gonad weight, and to the season of their capture (P < 0.05). A low prevalence (8.6%) and mean intensity (1.0) was detected in the muscle, probably related to the reduced ability of A. pegreffii to penetrate muscle. The risk of the presence of A. pegreffii in the muscle was fivefold higher in fish caught during the summer than during any other season. Susceptible members of the human population can minimize the risk of infection by avoiding the consumption of larger horse mackerel specimens during the summer.

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“…Allergens from the parasite are at present poorly characterized; many IgE-binding proteins remain unknown, and the clinical usefulness of different antigen sources (crude, heated, secretory, etc.) must be evaluated in the diverse diagnostic approaches (Baeza et al 2004;Moneo and Caballero 2002;Rodero et al 2004Rodero et al , 2005.…”

Section: Introductionmentioning

confidence: 99%

Sensitization to the fish parasite Anisakis simplex: clinical and laboratory aspects

Moneo¹,

Caballero

Rodríguez-Pérez

et al. 2007

Parasitol Res

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Hypersensitivity to Anisakis simplex is a worldwide medical problem. The parasite larvae die after freezing or cooking, but the tolerance of sensitized subjects to eating frozen fish remains a matter of controversy with contradictory findings. The aim of this study was to test if intolerance to properly cooked/frozen fish was due to the recognition of a particular allergen. Sixty-four patients with Anisakis simplex sensitization were studied by an IgE multiblot using simultaneously five different antigenic extracts. The antigens tested were a crude extract, excretory/secretory allergens, a heated extract, and two gradient ethanol precipitates of the crude extract. Intolerance was reported by 20% of the patients and was not related to the detection of any special allergen, nor to total or specific IgE levels. Intolerant patients only reported a higher frequency of digestive symptoms than the patients who tolerated fish ingestion. The most sensitive immunoblot antigen source was the 50-66% ethanol fraction of a crude extract (10x concentrated) that was found to be positive in 100% of the samples. Interestingly, 95% sensitivity in the IgE-immunoblot assay could be achieved using only two allergens, Ani s 1 and Ani s 4. Allergens from the dead larvae remain a problem for 20% of the sensitized subjects. The use of a fractionated and concentrated crude extract improved the sensitivity of the immunoblot assay.

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Enzyme-linked immunosorbent assay and Western blot antibody determination in sera from patients diagnosed with different helminthic infections with Anisakis simplex antigen purified by affinity chromatography

Abstract: An evaluation of the sensitivity and the specificity of the

Cited by 6 publications

References 26 publications

Western blot antibody determination in sera from patients diagnosed with Anisakis sensitization with different antigenic fractions of Anisakis simplex purified by affinity chromatography

Western blot antibody determination in sera from patients diagnosed with Anisakis sensitization with different antigenic fractions of Anisakis simplex purified by affinity chromatography

Epidemiology and molecular identification ofAnisakis pegreffii(Nematoda: Anisakidae) in the horse mackerelTrachurus trachurusfrom northern Morocco

Sensitization to the fish parasite Anisakis simplex: clinical and laboratory aspects

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