Testing of gastric contents for peanut proteins in a 13-year old anaphylaxis victim

Abstract

Background: We report the case of a 13-y female who went into anaphylactic shock following the ingestion of a meal suspected to be contaminated by peanuts. The teenager had a known sensitivity to peanuts, however, the restaurant claimed that no peanut products were used in the preparation of her meal. The gastric contents of the decedent were retained and tested for peanut proteins due to the possible legal liability of the proprietor.

Method: Using antibodies against peanut proteins (roasted and unroasted), we optimized a method to detect total soluble peanut proteins by Western-blot analysis in gastric contents. In addition, we validated two commercially available tests which were originally intended for detection of peanut proteins in food matrices to examine the same gastric sample. One was an enzyme-linked immunosorbent assay (ELISA) that utilized polyclonal antibodies against Ara h 1 (Tepnel Life Sciences). The other was a laminar-flow assay directed against Ara h 1, Ara h 2 and Ara h 3 (R-Biopharm). A positive food-based control was created by reducing bread and peanuts (1:1, w/w) with water (1:1, w/v) using a mortar and pestle. A food-based negative food control was created similar to the positive control, except the peanuts were omitted and the amount of bread was doubled.

Results: The Western-blot assay was sensitive down to 2.5ng/ml of total peanut protein. The laminar flow was the most rapid and least complex. The ELISA was the most analytically sensitive with a cut-off of 1ng/ml of Ara h 1 protein compared to the laminar flow which had a cut-off of 4ng/ml Ara h 1 equivalent. Both ELISA and laminar flow assays were able to detect peanut proteins in the food matrices and positive controls, and not in negative controls. No peanut related proteins were detected in the decedent's gastric sample. The gastric sample spiked with peanuts was reliably detectable.

Conclusion: The anaphylaxis patient had no peanut allergens detected in her gastric contents by any of the three methods employed. Both commercially available assays are easily adaptable for testing peanut allergens in the gastric contents as judged by the results of the immunoassays as well as the Western blot analysis. Due to the rising need for detecting peanut proteins in various heterogeneous and complex matrices, the use of appropriate controls should be also considered in these unique investigations.

Keywords: Anaphylaxis; Gastric Content; Peanuts; Soybean.