Carnés Jerónimo, Ferrer Angel, Huertas Angel Julio, Andreu Carmen, Larramendi Carlos H, Fernández-Caldas Enrique
Laboratorios LETI, S.L. Research Laboratories, Tres Cantos, Madrid, Spain.
Ann Allergy Asthma Immunol. 2007 Apr;98(4):349-54. doi: 10.1016/S1081-1206(10)60881-2.
Seafood plays an important role in human nutrition and is responsible for severe hypersensitivity reactions.
To evaluate how the cooking process may alter the in vivo and in vitro allergenicity of these extracts.
Raw and boiled extracts of shrimps and 2 types of lobsters were manufactured. Boiled extracts were prepared after the raw material was boiled for 15 minutes in phosphate-buffered saline. Raw and boiled extracts were homogenized and extracted for 4 hours. Afterward, the extracts were centrifuged, dialyzed, filtered, and freeze-dried. Seventy-eight patients were skin prick tested with these raw and boiled extracts. Specific IgE against the 6 extracts was measured by enzyme-linked immunosorbent assay (ELISA). Immunoblots and ELISA inhibition studies were performed with a pool of sera.
In vivo results showed that boiled extracts induced statistically significant larger wheals than raw extracts. More patients with positive results were also detected with boiled extracts. In vitro experiments by direct ELISA confirmed the in vivo results. The protein content in the boiled extracts decreased, and important differences were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cross-reactivity experiments showed that both types of extracts retained similar allergenic characteristics, even if the immunoblots revealed some differences in IgE binding.
More patients were identified using boiled extracts of shrimp and American and spiny lobsters than with raw extracts. The wheal sizes of the skin test reactions and specific IgE levels were also significantly greater using boiled extracts. The use of boiled extracts seems to be more effective in diagnosing seafood allergy.
海鲜在人类营养中起着重要作用，同时也会引发严重的过敏反应。
评估烹饪过程如何改变这些提取物的体内和体外致敏性。
制备虾和两种龙虾的生提取物和煮后提取物。将原料在磷酸盐缓冲盐水中煮沸15分钟后制备煮后提取物。将生提取物和煮后提取物匀浆并提取4小时。之后，对提取物进行离心、透析、过滤和冻干。用这些生提取物和煮后提取物对78名患者进行皮肤点刺试验。通过酶联免疫吸附测定（ELISA）测量针对这6种提取物的特异性IgE。用一组血清进行免疫印迹和ELISA抑制研究。
体内结果显示，煮后提取物诱导的风团在统计学上显著大于生提取物。用煮后提取物检测到的阳性结果患者也更多。直接ELISA的体外实验证实了体内结果。煮后提取物中的蛋白质含量降低，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测到重要差异。交叉反应实验表明，即使免疫印迹显示IgE结合存在一些差异，两种提取物仍保留相似的致敏特性。
与生提取物相比，使用虾、美国龙虾和多刺龙虾的煮后提取物能识别出更多患者。使用煮后提取物时，皮肤试验反应的风团大小和特异性IgE水平也显著更高。使用煮后提取物似乎在诊断海鲜过敏方面更有效。
