霜天蛾昆虫变应原多克隆抗体的制备及免疫特性分析

目的 制备兔抗霜天蛾变应原多克隆抗体,分析其与特异性过敏患者血清抗体成分的异同,为免疫筛选cDNA文库,制备基因重组霜天蛾变应原奠定基础.方法 应用霜天蛾变应原提取液免疫家兔获得霜天蛾变应原多克隆抗体,用酶联免疫吸附试验(ELISA)法测定混合免疫兔血清抗体效价:应用免疫印迹的方法对比分析免疫兔血清与特异性过敏患者血清抗体成分的异同.结果 混合免疫兔血清效价经ELISA检测,效价>1:10 000,免疫印迹检测兔抗血清免疫球蛋白G相似文献   

HCG酶联免疫试验诊断早期妊娠

我们采用绒毛膜促性腺激素(HCG)酶联免疫妊娠试验(双抗体夹心法)和免疫抑制凝集试验,对300例临床怀疑早孕的妇女及100例正常人(男50例,女50例)的晨尿进行了检测,并对其敏感性和特异性进行了测定。1.检测方法:HCG酶联免疫妊娠试验的     

青中年咳嗽变异性哮喘患者肺功能及变应原检测分析

目的分析青中年咳嗽变异性哮喘患者肺功能及变应原检测结果。方法前瞻性对比观察咳嗽变异性哮喘患者和正常人群FVC%、FEV1%、PEF%、MMEF%、FEF 25%、FEF 50%、FEF 75%、R Occ%指标,体外酶联免疫法检测患者血清变应原抗体。结果两组人群动态肺容积指标中,除FVC%、R Occ%无显著性差异(P0.05)外,其余FEV1%、PEF%、MMEF%、MEF75%、MEF50%、MEF25%指标中,观察组均低于对照组,两组存在显著性差异(P0.01);患者血清中发现变应原抗体的31例,占总数的44.93%。结论未经治疗的CVA患者大小气道气流均出现受阻现象,螨虫是诱导本地区青中年人群发生CVA最主要的变应原因素。     

霜天蛾昆虫变应原多克隆抗体的制备及免疫特性分析

目的制备兔抗霜天蛾变应原多克隆抗体,分析其与特异性过敏患者血清抗体成分的异同,为免疫筛选cDNA文库,制备基因重组霜天蛾变应原奠定基础。方法应用霜天蛾变应原提取液免疫家兔获得霜天蛾变应原多克隆抗体,用酶联免疫吸附试验(ELISA)法测定混合免疫兔血清抗体效价;应用免疫印迹的方法对比分析免疫兔血清与特异性过敏患者血清抗体成分的异同。结果混合免疫兔血清效价经ELISA检测,效价>1:10000,免疫印迹检测兔抗血清免疫球蛋白G(IgG)可以特异性识别霜天蛾10种蛋白组分,相对分子质量分别为100000、92000、79000、74000、66000、49000、43000、36000、25000和16000;患者血清IgG可以识别6种蛋白组分,相对分子质量分别为79000、74000、66000、49000、36000和25000。结论制备出的兔抗血清与特异性霜天蛾过敏患者血清特异性识别霜天蛾变应原的结果接近,比患者可以识别更多的蛋白条带,可以应用免疫兔血清作为筛选霜天蛾cDNA文库的探针及重组霜天蛾变应原表达蛋白的鉴定分析。     

变态反应疾病特异性诊断的规范化

变态反应疾病的特异性诊断应根据病史、体内试验结果、体外试验结果及变应原临床相关性综合分析进行判断，而不能单纯根据体内和，或体外试验阳性结果做出诊断。变态反应疾病特异性诊断必须规范化，应基于正确选择适应证及检测方法、检测方法的规范操作，以及对检测结果的临床意义给予正确解释。     

单克隆抗体在寄生虫病免疫诊断中的应用

单克隆抗体以其特异性高、效价高、易纯化等优点得到了广泛的应用,该文对单克隆抗体应用于乳胶凝集试验、间接血凝试验、免疫荧光检测、流式细胞仪检测、免疫组织化学、放射免疫测定法、免疫印迹技术、免疫酶染色技术和斑点免疫金银染色法、酶联免疫吸附试验、金免疫渗滤法、免疫层析技术和免疫色谱技术等12种免疫学诊断方法,及其在寄生虫病诊断中的应用作一综述.     

旋毛虫病免疫学诊断研究进展

旋毛虫病（trichinellosis）是一种危害严重的人畜共患寄生虫病,人因食入生的或不熟的含活幼虫囊包的肉类及其制品而感染.肌肉活检是该病确诊的依据,但是受摘取组织局限性的影响,不易检出幼虫,容易漏诊.免疫学诊断技术常常作为检测重要的辅助手段.该文概述了酶联免疫吸附试验、凝集试验、免疫酶染色试验、间接荧光抗体技术、快速免疫层析法、斑点免疫金渗滤法酶联免疫电转移印记技术等7种旋毛虫病免疫学诊断的抗体检测方法和1种抗原检测方法,并对早期、快速、准确诊断旋毛虫病的检测方法进行分析评价.     

变态反应学在中国

变态反应学是一门年轻的学科，在国外仅有百年，在我国则仅有50年的历史。我国变态反应学是从花粉症起步的。逐渐涉足到呼吸、皮肤、儿科等领域。经过全国同道的不懈努力，专业队伍不断壮大，目前我国各省市自治区都有了变态反应专业从业人员。共约2000余人。2001年成立了中华医学会变态反应学分会，目前有半数省份成立了变态反应学专业委员会，2007年《中华临床免疫和变态反应杂志》正式创刊。在我国，变应原的研究备受重视。早在20年前我国就进行过全国性花粉调查，继之又进行了真菌调查，在尘螨基础研究和应用研究方面也取得重要成果．宠物变态反应日益严重，食物变应原依次为牛奶、鸡蛋、坚果、油料作物、海鲜、水果、蔬菜和谷物。当前我国变态反应性疾病的特异性诊断方法仍为皮肤试验，包括皮内试验、点刺试验和斑贴试验。Immuno-CAP是最早进入我国的体外诊断方法．也是国内外公认的比较成熟的方法。近年来有多种方法进入我国市场，诸如免疫斑点、条带免疫印迹、放免、酶标、胶体金、化学发光、免疫捕获等方法。我们强调采用临床、体内、体外综合的特异性诊断方法．对变态反应性疾病必须采用防治结合。药物治疗与脱敏相结合的方法。目前，除尘螨外我国尚无商品化变应原，为解决学科发展的瓶颈问题。本文提出一些建议。     

尘螨变应原的免疫学研究进展

尘螨是一种变应原,能引起人类的过敏性疾病。三大类螨变应原的物埋化学特征、氨基酸顺序、酶的活性和B细胞抗原表位数已基本明了。重组螨变应原的抗原性之间存在差别。人体对螨变应原的免疫应答主要有变态反应和炎症反应学说,免疫应答中IgE产生与CD4~+T细胞(IL-4依赖)功能有关,且受MHC特异性限制。     

哮喘儿童体外过敏原检测及临床意义

目的:研究体外过敏原检测法在各年龄组哮喘儿童病因诊断中的临床意义。方法:应用UniCAP100系统的荧光酶联免疫方法,对106例临床确诊的哮喘患儿及86例无特异性疾病的对照组患儿进行吸入性变应原筛查(Phadiatop)和食物性变应原筛查(fx5),比较2组筛查的结果。哮喘组Phadiatop阳性者,选测尘螨、霉菌、猫及狗毛皮屑,树木和草花粉血清特异性IgE(SIgE)。fx5阳性,选测鸡蛋和牛奶SIgE。结果:变应原过筛总阳性率为67.92%,其中Phadiatop阳性率高于fx5,Phadiatop和fx5 2种检测的灵敏度为67.9%,特异度为88.4%。Phadiatop阳性率随年龄增加而增加,fx5则减少(P=0.001)。哮喘儿童变应原阳性率在不同性别、有无家族及有无本人特应性之间差异均无统计学意义(P>0.05)。结论:哮喘儿童2/3变应原阳性,婴幼儿期以食物性过敏原为主,吸入性过敏原随年龄而增加。吸入性变应原是哮喘儿童的主要病因。     

变应原突变体在支气管哮喘免疫治疗中的应用

特异性免疫治疗（SIT）是对明确了变应原的变态反应性疾病的一种有效的治疗方法。重组变应原及其衍生物的技术的进步大大提高了生产用于治疗变应性疾病的新型疫苗的能力。本文将就用定点突变技术获取的重组变应原突变体在支气管哮喘免疫治疗中的应用做一综述。     

Minimum prick test panel for adult patients with asthma and rhinitis in Ankara,Turkey

Objective: Determination of the number and type of allergens needed to be tested in epidemiological studies is important in order to identify most of the sensitized subjects with a cost-effective approach. This study aimed to investigate the minimum skin prick test panel for the identification of at least 95% of the sensitized subjects with symptoms of asthma and/or allergic rhinitis (AR) in Ankara, Turkey. Methods: Skin prick test results of 7492 patients who were referred to our outpatient clinic with clinical symptoms of asthma and/or AR between 1991 and 2005 were evaluated retrospectively. Seven allergens were tested in all and 13 allergens in 4202 patients. The allergen group needed for detection of 95% of the sensitized subjects was determined for both the 7 and 13 allergen panels. The study protocol was approved by the local ethics committee of Hacettepe University. Results: The atopy prevalences in the whole study population and in 4202 patients tested with the 13 allergen panel were calculated as 32.2% and 42.6%, respectively. Three allergens (Phleum pratense, Dermatophagoides pteronyssinus and Artemisia vulgaris) within the 7 allergen panel were adequate for the identification of at least 95% of the sensitized subjects. Olea europae was added to the previous three allergens when the 13 allergen panel was applied. Conclusion: Three to four allergens are sufficient for identification at least 95% of sensitized subjects with asthma and/or AR in Ankara, Turkey.     

IgE cross-reactivity between meadow fescue pollen and kiwi fruit in patients' sera with sensitivity to both extracts.

BACKGROUND: The presence of IgE reactivity to kiwi fruit and grass pollen allergens which could be caused by cross-reactivity has been detected in many patients with allergy. Proper identification of allergens as well as cross-reactive components is essential for understanding fruit- and pollen-associated hypersensitivity. METHODS: Using the sera from the polysensitized patients with specific IgE to grass pollen and kiwi fruit we tested reactivity to both allergen sources. IgE reactivity was exhibited in 8 serum samples by immunoblot. A serum pool formed by 8 individual sera was used for the investigation of IgE crossreactivity. SDS-PAGE immunoblot-inhibition assay was performed by preincubation of the sera with meadow fescue pollen, kiwi fruit extract, and isolated 24 kDa kiwi protein. To determine the allergens of kiwi fruit extract, we performed 2D PAGE immunoblot. In order to detect the crossreactive components between two allergen sources, a specific IgE for the 24 kDa kiwi allergen was purified. RESULTS: SDS-PAGE immunoblot meadow fescue pollen showed allergens ranging from 94 to 16 kDa, and kiwi fruit had 12 allergens ranging from 94 to 17 kDa. 2D-PAGE analysis revealed at least 15 spots in the kiwi extract and about 10 allergens. The most prominent allergen in 2D PAGE immunoblot was protein with 24 kDa and pI 9.4-9.5. Using an affinity-purified specific IgE we found that the 24 kDa kiwi allergen shared IgE-reactive epitopes with the meadow fescue group 4 and allergen about 36 kDa. Crossreactivity between isolated 24 kDa kiwi allergen and Fes p 4 was confirmed by anti-grass group 4 moab 2D8. CONCLUSION: Our findings showed that fescue meadow pollen cross-sensitize to kiwi fruits. A 24 kDa kiwi glycoprotein represent potential major allergen, which share common epitopes with Fes p 4 and 36 kDa meadow fescue allergen.     

武汉地区支气管哮喘患儿过敏原特异性IgE分析

目的：探讨武汉地区支气管哮喘儿童特异性过敏原分布及规律。方法：应用德国欧蒙公司生产过敏原特异性IgE检测试剂盒（中国组合）,采用免疫印迹法对216例武汉地区支气管哮喘儿童进行吸入性和食入性过敏原的血清学检测;并按年龄分为婴幼儿组（52例）和儿童组（164例）。对各组过敏原状况进行分析比较。结果：216例患儿中吸入性过敏原呈阳性162例（75％）。其中尘螨152例（70．3％）最高,屋尘89例（41．2％）、真菌组合78例（36．1％）分列其后;儿童组对吸入性过敏原及3种常见吸入性过敏原的阳性率比婴幼儿组高（P〈0．05）。食入性过敏原呈阳性52例（24％）,以虾蟹34例（15．7％）、海鱼26例（12％）、淡水鱼18例（8．3％）分居前3。不同年龄组阳性率差异无统计学意义。结论：过敏是哮喘发病重要因素,引起武汉地区儿童支气管哮喘的过敏原以吸入性过敏原为主并且阳性率随年龄增长而升高;尘螨是最重要的过敏原,食物性过敏原对哮喘的影响亦需重视。     

Biology of tree pollen allergens

More than 25% of the population suffer from type I allergy. Pollens from trees of the Fagales, Oleaceae, and Cupressaceae belong to the most potent and frequent allergen sources. During the past 15 years, the nature of the most important allergens has been identified by molecular biological techniques, and recombinant allergens equivalent to the natural allergens have been produced. These advances provide insight into the biological functions of important allergens and allow the development of novel forms of diagnosis and therapy. In this review, we focus on Fagales allergens to illustrate the impact of recombinant allergens on diagnosis and therapy. We discuss structural similarities as a molecular basis for crossreactivities and develop diagnostic concepts by using speciesspecific marker allergens as well as highly cross-reactive allergens. The identification of the allergen recognition profiles of patients with recombinant allergens allows a more precise selection of patients for available forms of allergy treatment. Moreover, we describe novel recombinant allergen-based forms of specific immunotherapy.     

Hidden allergic factors in the etiology of asthma

Increasing evidence from case control surveys, population studies and allergen avoidance studies suggest inhalant allergy plays an important role in the etiology of asthma. Recent studies in hospital emergency rooms have compared the prevalence of serum IgE antibodies to common allergens (mite, cat, cockroach, rye grass and ragweed pollen) in patients admitted with acute asthma attacks and in unselected age-matched control subjects. These studies, carried out in central Virginia and northern California, showed a highly increased prevalence of IgE antibodies to inhaled allergens among asthmatic patients, and suggest that the development of allergen specific IgE antibody responses is a major risk factor for emergency room admission with asthma. Presentation at the emergency room appeared to be related to patients' exposure to specific allergens: in central Virginia, in the fall, dust mite was the predominant allergy, whereas in northern California, in May-June, most asthmatic patients (greater than 90 percent) were allergic to rye grass. New immunoassay technology, based on the use of monoclonl antibodies, has been developed to measure the quantities of "indoor" allergens (mite, cat, cockroach) in asthmatic patients' houses. It is now possible to propose tentative levels of mite allergens which should be considered both as a risk for IgE antibody sensitization (2micrograms allergen/g dust) and as a risk for acute asthma attacks (10micrograms allergen/g dust). Future management of asthma will require analysis of indoor allergens and the development of efficient allergen avoidance procedures. Further research is necessary to investigate the relationship between airborne allergen levels, particle size and the precipitation of asthma attacks and also to investigate immunologic mechanisms which may cause bronchial hyperreactivity.     

Les allergènes recombinants : leur apport à l’allergologie en 2006

Advances in molecular biology techniques have led to the production of recombinant allergens, about thirty of them now being available for measurements (DIAGNOSIS?) in vitro. These recombinant allergens correspond to a precise molecular variant of a natural allergen, and their biological activity has to be evaluated in comparison with the corresponding natural allergen. The advantages of recombinant allergens are essentially the creation of allergenic preparations having constant pharmaceutic properties, which allows determination of specific IgE directed against different molecular components of an allergenic source, for example, pollen, mites, etc. The main consequences of these biological advances are the following: evaluation of sensitivities to allergen molecules in different populations (molecular epidemiology), improvement of extracts used for diagnosis by selection of the most pertinent allergenic sources and in quantifying their major allergen content, definition of the spectrum of recognition of specific IgE vis-à-vis different molecular components (spectrotype), quantitative evaluation of IgE responses, establishing the molecular basis of cross-reactions between different inhaled allergens, between different food allergens, and between inhaled allergens and food allergens. As regards allergy practice, this new diagnostic tool can lead to better interpretation of polysensitivities, observed by skin tests and in vitro tests. Some examples of particular clinical cases associated with specific sensitivities vis-à-vis certain recombinant allergens will be presented.     

Faut-il évaluer et doser la charge allergénique environnementale ?

Assessment of the indoor allergen load encountered by patients with respiratory allergy symptoms is essential in order to obtain objective evidence of the allergens present in the indoor environment. It is equally essential to justify application of efforts to eliminate the responsible allergen sources (mites, domestic animals, molds, etc.). The methods used to measure indoor allergens can be grouped into (1) quantitative immunoassays (ELISA using monoclonal antibodies specific for purified native allergens or recombinant allergens) and (2) home-based tests. The latter include the Acarex test (semiquantitative assay for guanine in mite feces) and semiquantitative immunoassays. Quantitative tests are used mainly for epidemiologic studies whereas the results of home-based tests are considered in the management of allergic patients.     

Allergènes alimentaires croisant avec les allergènes des pollens

The range of pollen-food cross-reactions has increased over the past decade, the clinical pictures are more clear with respect to the allergens concerned, and the molecular basis for some of them have been determined. Diagnostic methods include skin tests, assays for specific IgE, and open and double blind oral provocation tests. Investigation of allergic cross-reactions, first based solely on immunological inhibition techniques using natural allergen extracts, have benefited from molecular biology. Many allergens homologous with pollen allergens have been sequenced and the three dimensional structure of Pru av 1 has been determined, allowing studies on a sub-molecular scale. Allergen cross-reactions between pollen and food, for which the clinical relevance is well established, involve allergens of the Bet v 1 and Bet v 2 families. These allergens are present in numerous edible fruits and vegetables. Identity with the Bet v 1 sequence varies from 38 to 67%, being closest for the profilins (70-80%). Cross sensitization with Bet v 1 and profilins, although particularly frequent, may be silent clinically. Other candidate molecules involved in cross-reactions between pollen and food allergens are Bet v 6, a minor birch allergen, the lipotransferases found in some of the compositae, and the 1,3-β-glucanases corresponding to a major olive allergen. The significance of the detection of specific IgE directed against carbohydrate determinants remains to be investigated. A major problem for the clinician is the absence of clinical significance of cross-reactivity that has been demonstrated in vitro and in vivo.     

Laboratory tests for diagnosis of food allergy: advantages,disadvantages and future perspectives

Numerous biological tests point to the diagnosis of food sensitization: detection of specific IgEs by Rast techniques, multi-detection assays, immunoblotting, screening of basophil activation (BAT or FAST), assays for leukotriene LTC4 release (CAST), measurement of plasma histamine, serum tryptase, serum ECP, urinary EDN, completed by mannitol-lactulose test evaluating intestinal permeability, assay of fecal IgEs, Rast for specific IgG4. Primary screening for anti-food IgEs by multi-detection assays seeks justification from insufficient clinical data and false positive tests are common in patients sensitized to pollens or latex, on account of in vitro cross reactivities (CR). Multiple CR explain positive Rast to vegetal food allergens in such patients. Biological tests should not be performed as the first line of diagnosis. In vivo sensitisation is assessed by positive prick-tests, demonstrating the bivalence of allergens, as well as the affinity of specific IgEs, two conditions necessary to bridge membrane bound specific IgEs, leading to the release of mediators. Prick-tests are closer to clinical symptoms than biological tests. However, the diagnosis of food allergy is based on standardised oral challenges. Exceptions are high levels of specific IgEs to egg (> 6 kUl/l), peanut (> 15 kUl/l), fish (> 20 kUl/l) and milk (> 32 kUl/l), reaching a 95% predictive positive value. Rast inhibition tests are useful to identify masked allergens in foods. Research developments will have impact on the development of new diagnostic tools: allergen mixes reinforcing a food extract by associated recombinant major allergens, multiple combination of recombinant allergens (chips) or tests with synthetic epitopes aimed a the prediction of recovery. Laboratory tests take place in the decision free for the diagnosis for the food allergy and the follow-up of the levels specific IgEs is a tool to assess outcome and contributes to predict recovery or persistent allergy. Up to now the significance of positive laboratory tests showing the implication of IgEs is at the crossroads of the allergist's and biologist's expertise.