Bugajska-Schretter et al. (2000) [1314] reported the symptoms of 13 patients allergic to fish (10 adults, three children), all of whom had IgE antibodies binding carp parvalbumin. All patients were allergic to more than one fish species and experienced at least one of the typical clinical symptoms (dermatitis, urticaria, angioedema, diarrhoea, asthma, anaphylactic reaction) after contact with fish proteins.

Bugajska-Schretter et al. (1998) [1318] similarly report the symptoms of 30 fish allergic patients as dermatitis, urticaria, angioedema, diarrhea, asthma, or anaphylactic reaction after ingestion, inhalation, or skin contact with fish proteins. Although not all symptoms were not specifically associated with carp, the 28 sera tested from these patients showed IgE binding to carp parvalbumin. Data on 4 representative patients noted that 2/4 had shown clinical symptoms after carp ingestion of urticaria and urticaria plus asthma .

Bugajska-Schretter et al. 2000 [1314] tested sera from 13 patients allergic to fish (10 adults, three children) with a positive case history of type I allergy to fish.

Bugajska-Schretter et al. (1998) [1318] tested sera from 30 patients allergic to fish with a positive case history of type I allergy to fish. 28 were tested with carp parvalbumin by immunoblotting.

Bugajska-Schretter et al. (1998) [1318] report positive RASTs, class 2-4, with all 30 patients with cod extracts and class 1-4 with salmon and tuna (only 29 tested with tuna). The degree of inhibition of IgE binding to tuna, salmon, perch, carp and eel obtained after preadsorption of the four sera with cod extract was determined by gamma counting giving 43%, 93%, 84% and 73% for carp. 

Bugajska-Schretter et al. (1998) [1318] and Bugajska-Schretter et al. 2000 [1314] blotted proteins separated by SDS-PAGE or IEF onto nitrocellulose membranes (Nitrocell, Pharmacia Biotech) or polyvinylidenedifluoride membranes (Immobilon PVDF, Millipore, Bedford, Massachusetts, USA) by the method of Towbin et al. 1979 [1069]. Nitrocellulose strips and PVDF membranes containing blotted proteins were blocked at room temperature in PBS, pH 7.5, containing 0.5% (v/v) Tween 20. Strips were then probed overnight at 4°C with sera, diluted 1:10 (v/v) in the same buffer. Bound IgE antibodies were detected with ¹²⁵I-labelled anti-human IgE antibodies (Pharmacia) and visualised by autoradiography.

Bugajska-Schretter et al. 2000 [1314] report that binding of IgE from 4 sera from fish allergic patients to carp parvalbumin is reduced 59%, 51%, 79%, and 27% on removal of calcium. Preincubation of sera with purified pI 4.7 form of carp parvalbumin strongly inhibited IgE binding to the other isoforms and to extracts of cod fish, tuna, and salmon, absorbing, on average, 83% of fish-specific IgE antibodies. Similarly, Preabsorbtion of sera from fish allergic patients by purified carp parvalbumin either abolished or greatly reduced IgE binding to nitrocellulose-blotted extracts of carp, cod, eel, perch or salmon (Swoboda et al 2002 [1787]).

Bugajska-Schretter et al. (1998) [1318] reported that all 28 sera contained IgE binding a protein in cod and carp extracts at 12 kDa (parvalbumin). 7 sera also bound other minor allergens in cod extract of 200, 100, 80, 66, 35, 30, 25, 20, and 15 kDa. Periodate treatment of the blotted extracts strongly reduced IgE binding to parvalbumin with the 3 sera tested, suggesting that either carbohydrate or free cysteine residues are involved in IgE binding. Calcium depletion with EGTA strongly reduced binding to parvalbumin but not to minor allergens with 9/14 sera tested. Reduction did not change IgE binding. The degree of inhibition of IgE binding to tuna, salmon, perch, carp and eel obtained after preadsorption of the four sera with cod extract was determined by gamma counting giving 43%, 93%, 84% and 73% for carp.