本文選題：交叉過敏 + 塵螨��； 參考：《河北醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:過敏反應(yīng)主要由血清中免疫球蛋白E(Ig E)介導(dǎo),通過效應(yīng)細(xì)胞釋放生物活性介質(zhì)而產(chǎn)生各種臨床表現(xiàn),如蕁麻疹、血管神經(jīng)性水腫等。有時(shí),即使是微量蛋白質(zhì)也可引起敏感個(gè)體出現(xiàn)致命的反應(yīng)。隨著對(duì)過敏性疾病的不斷研究,交叉過敏反應(yīng)也逐漸引起人們的重視。所謂交叉過敏就是當(dāng)兩種蛋白質(zhì)在序列或結(jié)構(gòu)上高度同源時(shí),出現(xiàn)相同的抗原決定簇的幾率增高,易出現(xiàn)交叉反應(yīng)。檢測(cè)屋塵螨與雞蛋白之間是否存在交叉過敏、交叉過敏發(fā)生的概率以及發(fā)生交叉反應(yīng)的蛋白種類;屋塵螨及雞蛋白市售蛋白分子組分;比較免疫印跡法與狹縫免疫印跡法(Slot-blot)符合概率�；谀壳拔輭m螨與雞蛋白過敏的發(fā)病率高,診斷及防治難的特點(diǎn),本研究成果為屋塵螨與雞蛋白過敏提供了可行且可靠的個(gè)體化治療方法,并且使臨床醫(yī)師能更好的指導(dǎo)過敏患者正確飲食及生活習(xí)慣。方法:1流行病學(xué)調(diào)查及分析:對(duì)2015.01-2016.10就診于河北醫(yī)科大學(xué)第四醫(yī)院皮膚科進(jìn)行血清特異性過敏原檢測(cè)的患者進(jìn)行流行病學(xué)調(diào)查,并結(jié)合sIgE檢測(cè)結(jié)果進(jìn)行回顧性分析。2選取塵螨特異性sIgE(specific Ig E,sIgE)與總Ig E(total Ig E,t Ig E)同時(shí)陽性血清進(jìn)行狹縫免疫印跡實(shí)驗(yàn)(Slot-blot),對(duì)比Slot-blot檢測(cè)結(jié)果與艾康生物技術(shù)有限公司銷售的免疫印跡法結(jié)果的一致性。3采用Slot-blot法進(jìn)行交叉實(shí)驗(yàn),采用雞蛋白sIgE與t Ig E同時(shí)陽性血清與屋塵螨抗原進(jìn)行反應(yīng),檢測(cè)塵螨與雞蛋白是否存在交叉反應(yīng)及交叉概率。4采用Slot-blot進(jìn)行抑制實(shí)驗(yàn),先將雞蛋白與屋塵螨sIgE陽性血清進(jìn)行反應(yīng),然后再與包被的屋塵螨抗原反應(yīng),檢測(cè)屋塵螨與雞蛋白交叉抑制率和有效抑制濃度。5將屋塵螨及雞蛋白粗提取液進(jìn)行聚丙烯酰氨凝膠電泳(SDS-PAGE),檢測(cè)各蛋白組分的完整性、分子量大小及所占比例。6對(duì)屋塵螨和雞蛋白粗提取液進(jìn)行免疫印跡實(shí)驗(yàn)(Western bolt,WB),選取屋塵螨sIgE與t Ig E同時(shí)陽性血清和雞蛋白血sIgE與t Ig E同時(shí)陽性血清各10例為探針,檢測(cè)各組分的抗原性。結(jié)果:1流行病學(xué)調(diào)查1.1本研究對(duì)2015.01-2016.10就診于河北醫(yī)科大學(xué)第四醫(yī)院皮膚科的診斷為過敏反應(yīng)相關(guān)性疾病的患者血清sIgE檢測(cè)結(jié)果進(jìn)行了回顧性分析。病例總數(shù)為5186例,男3100例,女2086例,年齡31.72±1.12歲,病程1周-數(shù)年,病種以蕁麻疹、濕疹及皮炎為主。其中t Ig E陽性為3828例,塵螨sIgE陽性患者1223例,雞蛋白sIgE陽性患者822例,其中塵螨與雞蛋白sIgE同時(shí)陽性病例達(dá)419例,塵螨與雞蛋白sIgE同時(shí)陽性病例占?jí)m螨sIgE陽性的病例的34%。1.2過敏原sIgE陽性病例的分布情況:塵螨1223(31.9%),雞蛋白822(21.5%),牛奶430(11.2%),牛羊肉344(9%),小麥223(5.8%),貓狗毛300(7.8%),霉菌172(4.5%),矮豚草166(4.3%),樹組合114(3%),蟑螂109(2.8%),凬草96(2.5%),腰果花生159(4.2%),魚蝦蟹152(4%),芒果150(3.9%)。2免疫印跡法(艾康生物有限公司)與Slot-blot法實(shí)驗(yàn)結(jié)果對(duì)比分析60例塵螨sIgE陽性(免疫印跡法)的血清中,56例在Slot-blot實(shí)驗(yàn)中顯示陽性,一致性概率達(dá)95%。3屋塵螨抗原與雞蛋白陽性血清交叉過敏反應(yīng)概率(Slot-blot法)將包埋于PVDF膜上的屋塵螨抗原與雞蛋白sIgE陽性血清反應(yīng),隨機(jī)檢測(cè)60例,30例出現(xiàn)陽性反應(yīng),其出現(xiàn)交叉反應(yīng)概率為50%。4雞蛋白對(duì)屋塵螨抗原抗體反應(yīng)的交叉抑制率的檢測(cè)雞蛋白對(duì)屋塵螨抗原抗體反應(yīng)有明顯的抑制作用,且呈濃度依賴性,當(dāng)雞蛋白干預(yù)抗原濃度為100μg/ml時(shí),達(dá)到最大抑制率,為93%。5屋塵螨提取液與雞蛋白提取液中各蛋白組分分析5.1屋塵螨列可見3條主要條帶,分別是57kd,50kd,16kd,各組分所占比例依次是55.5%,43.4%,1.3%,其中57kd的蛋白所占比例最大。5.2雞蛋白列可見4條主要條帶,分別是75kd,44.5kd,28kd,14.3kd,各組分所占比例依次是19.6%,66%,7.7%,6.6%,其中所占比例最大的是44.5kd的條帶。6屋塵螨與雞蛋白免疫抗原性分析(WB檢測(cè))6.1 10例塵螨sIgE陽性患者的血清池為探針,50kd、57kd處可見明顯陽性反應(yīng)條帶,考慮是Derp10(原肌凝蛋白)和Derp4(α-淀粉酶)。6.2 10例雞蛋白sIgE陽性的患者血清池為探針。主要陽性反應(yīng)條帶有3條,分別是14.3kd、44.5kd、75kd,考慮分別為溶菌酶(Lys)、卵白蛋白(OVA)和卵轉(zhuǎn)鐵蛋白(OVT)。結(jié)論:1就診于我院的過敏反應(yīng)相關(guān)疾病的患者,主要過敏原是塵螨組合和雞蛋白。2屋塵螨主要過敏原為Derp10(原肌凝蛋白)和Derp4(α-淀粉酶);雞蛋白主要過敏原為溶菌酶(Lys)、卵白蛋白(OVA)和卵轉(zhuǎn)鐵蛋白(OVT)。3屋塵螨與雞蛋白兩種抗原之間存在相似的抗原表位,又存在各自特異性表位。4屋塵螨與雞蛋白兩種蛋白質(zhì)由于存在相似抗原表位,而導(dǎo)致交叉過敏反應(yīng)的發(fā)生。5雞蛋白對(duì)屋塵螨的抗原抗體反應(yīng)有交叉抑制作用,且抑制程度有濃度依賴性,當(dāng)雞蛋白濃度為100μg/ml時(shí)可達(dá)最大抑制,其抑制率為93%。
[Abstract]:Objective: Anaphylaxis is mainly mediated by immunoglobulin E (Ig E) in the serum, and produces various clinical manifestations, such as urticaria, vascular neuroedema, and so on. Sometimes, even trace proteins can cause a fatal response to sensitive individuals. Anaphylaxis is also attracting more and more attention. The so-called cross allergy is the increase in the probability of the same antigen determinant when two proteins are highly homologous in sequence or structure. Cross reaction is easy to occur. Protein species, house dust mite and chicken protein Market protein components; comparison between Western blot and slit immunoblotting (Slot-blot). Based on the high incidence of house dust mites and egg white hypersensitivity and difficult diagnosis and prevention, this study provides a feasible and reliable individualized treatment for house dust mites and egg white allergies. Methods: and enable clinicians to better guide the correct diet and living habits of allergic patients. Methods: 1 epidemiological investigation and analysis: 2015.01-2016.10 in the Department of Dermatology, the Department of Dermatology, the fourth hospital of Hebei Medical University, carried out the epidemiological investigation of the patients with specific allergen detection in the serum, and reviewed the results of the test in combination with the results of the test. Sex analysis.2 selected dust mites specific sIgE (specific Ig E, sIgE) and total Ig E (total Ig E, t) at the same time to carry out the slit immunoblotting experiment. The positive sera of white sIgE and t Ig E reacted with the house dust mite antigen to detect the cross reaction between the dust mite and the chicken protein and the cross probability of the chicken protein,.4 using Slot-blot to inhibit the test. First, the chicken protein was reacted with the positive serum of the house dust mite sIgE, and then it was reacted with the antigen of the house dust mites, and the cross inhibition of the house dust mites and chicken protein was detected. .5 and SDS-PAGE were carried out to extract the extract of house dust mite and egg white, and the integrality, molecular weight and proportion of.6 were detected by.6 in the immunoblotting test (Western bolt, WB) of house dust mite and egg white extract, and the positive serum of sIgE and t Ig E were selected as the positive serum. 10 cases of egg white blood sIgE and t Ig E positive serum were used as probes to detect the antigenicity of each component. Results: 1 epidemiological investigation was conducted in 1.1 studies on the retrospective analysis of the results of serum sIgE detection in patients diagnosed as allergic reaction related diseases in Department of Dermatology, Hebei Medical University, Hebei Medical University. The total number was 5186 cases, 3100 males and 2086 females, with age 31.72 + 1.12 years, and the course of disease was 1 weeks for 1 weeks. The disease species were urticaria, eczema and dermatitis. Among them, t Ig E positive was 3828 cases, dust mites sIgE positive patients were 1223, and egg white sIgE positive patients were 822 cases, including 419 cases of dust mite and chicken protein sIgE positive cases, dust mites and egg white sIgE simultaneously Yang The distribution of 34%.1.2 allergen positive cases in the cases of sIgE positive cases of dust mites were 1223 (31.9%), egg white 822 (21.5%), milk 430 (11.2%), beef and sheep meat 344 (9%), wheat 223 (5.8%), cat dog hair 300 (7.8%), fungus 172 (4.5%), dwarf ragweed, cockroach herb, cashew peanut 152 (4%), mango 150 (3.9%).2 immunoblotting (ikang bio limited company) and Slot-blot test results were compared and analyzed in the serum of 60 cases of sIgE positive (immunoblotting). 56 cases were positive in the Slot-blot test, and the consistency probability reached the probability of cross allergic reaction between 95%.3 house dust mite antigen and egg white positive serum (Slot-bl). Ot method) the reaction of house dust mite antigen embedded in PVDF membrane and positive serum of egg white sIgE was detected in 60 cases randomly and 30 cases were positive. The cross reaction probability of cross reaction was the inhibition rate of cross reaction of 50%.4 chicken protein to house dust mite antigen antibody reaction, and the chicken protein had obvious inhibition effect on the antibody reaction of house dust mite antigen. Degree dependence, when the concentration of chicken protein intervention antigen was 100 g/ml, the maximum inhibition rate was reached. 3 main bands were visible for the analysis of 5.1 house dust mite in the 93%.5 house dust mite extract and the chicken protein extract, respectively, 57kd, 50kd, 16kd, respectively. The proportion of each group was 55.5%, 43.4%, 1.3%, of which the 57kd protein accounted for the largest proportion. 5.2 egg white shows 4 main bands, which are 75kd, 44.5kd, 28kD, 14.3kd. The proportion of each group is 19.6%, 66%, 7.7%, 6.6%. The largest proportion is the 44.5kd strip.6 house dust mite and the chicken protein immune antigenicity analysis (WB test) 6.110 cases of dust mite sIgE positive patients' serum pool as the probe, 50kd, 57kd can be visible obvious The positive reaction strip was considered as the probe of the serum pool of Derp10 (primary myoprotein) and Derp4 (alpha amylase).6.2 10 cases of chicken protein sIgE positive. The main positive reaction strips were 3, respectively 14.3kd, 44.5kd, 75kd, respectively, which were considered to be lysozyme (Lys), egg white (OVA) and oval transferrin (OVT). Conclusion: 1 is the allergic reaction in our hospital. The main allergen of the related disease is the combination of dust mites and the main allergy of.2 house dust mites: Derp10 (procoagulin) and Derp4 (alpha amylase); the main allergy of egg white is lysozyme (Lys). There are similar epitopes between the two antigens of the ovalbumin (OVA) and the oval transferrin (OVT).3 house dust mite and egg white. The two proteins,.4 house dust mite and egg white in their specific epitopes, have similar antigen epitopes, which lead to cross anaphylaxis,.5 chicken protein has a cross inhibition effect on the antigen antibody response of house dust mite, and the inhibition degree is concentration dependent. When the chicken protein concentration is 100 g/ml, the maximum inhibition is reached and the inhibition rate is 93%..
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R593.1;R446.6

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