發(fā)布時(shí)間：2018-04-15 21:19

  本文選題：刀額新對(duì)蝦 + 過(guò)敏原　； 參考：《江南大學(xué)》2008年碩士論文

【摘要】： 食物過(guò)敏作為一類食源性疾病已引起了人們的普遍關(guān)注。過(guò)敏性疾病已被世界衛(wèi)生組織列為二十一世紀(jì)重點(diǎn)防沫的三大類疾病之一,是當(dāng)前世界性的重大衛(wèi)生學(xué)問(wèn)題。食物過(guò)敏作為一類常見的過(guò)敏性疾病,引起的臨床癥狀主要有頭暈、頭痛、胸悶、惡心、嘔吐、皮膚痙癢等,嚴(yán)重時(shí)還會(huì)休克甚至死亡。因此,開展食物(特別是海產(chǎn)品)的過(guò)敏原研究對(duì)保護(hù)人們生命健康,提高人民生活質(zhì)量具有巨大的經(jīng)濟(jì)效益和社會(huì)效益。 蝦蟹等甲殼類動(dòng)物及其制品是人類優(yōu)質(zhì)動(dòng)物蛋白的來(lái)源之一,也是八大類容易引起過(guò)敏的食物之一。隨著國(guó)際貿(mào)易的發(fā)展,其在食品中的應(yīng)用越來(lái)越廣泛,食用蝦引起過(guò)敏的潛在危害更值得重視。本論文選用我國(guó)華南地區(qū)常見的食用蝦種,刀額新對(duì)蝦(Metapenaeus ensis俗名基圍蝦),作為研究對(duì)象,主要從蝦過(guò)敏原蛋白的分離純化、純化產(chǎn)物的免疫學(xué)鑒定等方面進(jìn)行研究,得到如下結(jié)論: 首先,應(yīng)用SDS-PAGE技術(shù)對(duì)粗提蛋白液進(jìn)行初步分離,SDS-PAGE圖譜顯示刀額新對(duì)蝦粗提蛋白約有20條可辨蛋白帶,分子量在12.1 KDa～105.6 KDa之間。利用12例蝦過(guò)敏患者血清通過(guò)免疫印跡(western-blotting)分析,初步鑒定出刀額新對(duì)蝦粗提液中有7條過(guò)敏蛋白條帶,其分子量分別為36 KDa、46.2 KDa、60 KDa、64.8 KDa、68 KDa、78 KDa和88.5 KDa。另外還有21.6 KDa的蛋白也被個(gè)別患者的血清識(shí)別,其中4條過(guò)敏蛋白條帶反應(yīng)率高,為主要致敏組分,分子量分別是36 KDa、46.2KDa、60 KDa和68 KDa,這與國(guó)內(nèi)外其他人的研究結(jié)果有一定差異,這可能與蝦的種類不同、各地區(qū)蝦類生長(zhǎng)環(huán)境的不一致性以及過(guò)敏患者人群的個(gè)體差異有關(guān)。 其次,通過(guò)硫酸銨分段鹽析、DEAE-52離子交換層析和Sephadex G-100凝膠柱層析等方法逐步純化了蝦過(guò)敏蛋白。并通過(guò)免疫印跡(western-blotting)、斑點(diǎn)酶聯(lián)免疫(dot-ELISA)、和酶聯(lián)免疫(ELISA)間接法對(duì)已經(jīng)純化的蝦過(guò)敏蛋白進(jìn)行了分析和鑒定。具有很高的反應(yīng)性。 本研究初步純化、鑒定出刀額新對(duì)蝦主要過(guò)敏原蛋白,為進(jìn)一步研究蝦類過(guò)敏原的結(jié)構(gòu)與功能奠定了一定的理論基礎(chǔ),其在ELISA檢測(cè)中作為特異性抗原與蝦過(guò)敏癥患者血清中特異性IgE有良好的反應(yīng)性,也為蝦類食物過(guò)敏反應(yīng)疾病的診斷和進(jìn)一步研制脫敏試劑奠定了良好的基礎(chǔ)。
[Abstract]:Food allergy, as a kind of foodborne disease, has attracted people's attention.Allergic diseases have been listed by the World Health Organization as one of the three major anti-foaming diseases in the 21 century.Food allergy as a common allergic disease, the main clinical symptoms caused by dizziness, headache, chest tightness, nausea, vomiting, skin spasmolysis and itching, severe shock and even death.Therefore, the development of food (especially seafood) allergen research to protect people's life and health, improve people's quality of life has great economic and social benefits.Crustaceans such as shrimps and crabs and their products are one of the sources of human high quality animal protein and one of the eight kinds of food which can easily cause allergies.With the development of international trade, its application in food is more and more extensive.In this paper, the common edible shrimp species in South China, Metapenaeus ensis, as the research object, were studied from the isolation and purification of the allergen protein and the immunological identification of the purified products.The following conclusions have been drawn:Firstly, the SDS-PAGE analysis of crude protein extract by SDS-PAGE showed that there were about 20 discernible protein bands with molecular weight between 12.1 KDa~105.6 KDa and 12.1 KDa~105.6 KDa.Using Western blotting analysis of the sera of 12 shrimp allergy patients, we preliminarily identified 7 bands of allergy protein in the crude extract of Penaeus Monodon, the molecular weight of which was 36 K Dax 46.2 K Da1 60 K Da4. 8 K DaDX 68 KDaf78 KDa and 88 5 K Da. respectively.In addition, the protein of 21.6 KDa was also recognized by the serum of individual patients. The reaction rate of four allergic protein bands was high, and the molecular weight was 36KDa46.2KDa-60 KDa and 68kDa. the results were different from those of other people at home and abroad.This may be related to the different species of shrimp, the inconsistency of shrimp growth environment in different regions and the individual difference of allergy patients.Secondly, shrimp allergy protein was purified by ammonium sulfate fractionated DEAE-52 ion exchange chromatography and Sephadex G-100 gel column chromatography.The purified shrimp allergy proteins were analyzed and identified by Western blotting, dot-ELISAA and Elisa indirect methods.Be highly reactive.In this study, the main allergen proteins were identified by purification, which laid a theoretical foundation for further study on the structure and function of shrimp allergens.As a specific antigen in the detection of ELISA, it has good reactivity with specific IgE in the sera of shrimp hypersensitivity patients. It also lays a good foundation for the diagnosis of shrimp food allergy disease and the further development of desensitizing reagent.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)期刊論文 前3條

1 吳海明;胡志和;王超;;免疫印跡法對(duì)4種海蝦主要過(guò)敏原的鑒定[J];食品科學(xué);2010年16期

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相關(guān)碩士學(xué)位論文 前1條

1 阮韋偉;糖基修飾改性對(duì)擬穴青蟹過(guò)敏原性質(zhì)的影響[D];集美大學(xué);2012年

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本文編號(hào)：1755818