Allergen micro-array detection of specific IgE-reactivity in Chinese allergy patients

Background  Allergen micro-arrays are powerful tools for screening of serum IgE-reactivity. In this study allergen micro-arrays were used to identify dominating IgE-binding allergens and cross-reactivity patterns among selected Chinese allergy patients.

Methods  The study was conducted using patient sera from the cities of Guangzhou, Nanjing, Chengdu and Shenyang. In total 100 sera with Dermatophagoides pteronyssinus (Der p) specific IgE-levels higher than 50 kU/L were selected for testing against 103 individual allergens.

Results  Among 100 selected patients, 95% showed IgE-reactivity towards house-dust mite allergens Dermatophagoides farinae (Der f) 1, Der f 2 and Der p 2 and 94% were IgE positive against Der p 1, and 60% of sera contained IgE reacting against allergen Euroglyphus maynei (Eur m) 2. IgE against cat allergen, Felisdomesticus (Fel d) 1, was seen in 20%. Only 2% showed specific IgE-reactivity to Der p 10, a panallergen belonging to the tropomyosin family. Serum IgE-reactivity towards other allergens was in general low. IgE-reactivity against pollen allergens showed geographic differences.

Conclusions  This study clearly confirms that group 1 and group 2 are major allergens of house dust mites. These selected house-dust mite allergy patients are close to being mono-sensitized. Der p 10 is not an important allergen for cross-reactivity. Specific IgE-sensitization towards pollen allergens is low in southern China compared to other regions. The prevalence of food and stinging insect allergens known to give rise to IgE-mediated cross-reactivity is 2% or less.

     

变态反应性疾病患者螨和蟑螂过敏原体外检测分析

目的探讨变态反应性疾病患者血清中螨和蟑螂过敏原特异性免疫球蛋白E(SIgE)的情况以及它们的相互关系分析。方法采用荧光酶联免疫吸附法,对我院2004年10月至2005年10月就诊于呼吸科和变态反应科门诊的199例患者,检测其血清中螨和蟑螂的SIgE。结果在变态反应性疾病的患者中,对于螨类过敏的,包括屋尘螨(d1)、粉尘螨(d2)和热带无爪螨(d201)均为儿童组比成年组的阳性率高,同时具有较高的强阳性率;对于热带无爪螨过敏的患者,一般都是在4级以下,极个别具有强阳性且均为儿童;对于蟑螂过敏的,所有患者均在4级以下;对于同时检测尘螨和蟑螂的患者,其血清中的SIgE值均为各螨类显著高于蟑螂的值(P<0.01)。结论尘螨和蟑螂均为变态反应性疾病的重要过敏原,尤其是尘螨;对于尘螨和蟑螂同时过敏的患者,尘螨的过敏程度均比蟑螂高。     

Indoor Allergen Levels and Household Distributions in Nine Cities Across China

Objective Chinese allergic subjects have high levels of sensitization to house dust mite (HDM) and other indoor allergens. This study quantifies common indoor allergen levels in Chinese households.
Methods Dust samples were collected from nine cities. Major allergens Der p 1 and Der f 1 from Dermatophagoides pteronyssinus and D. farinae, and specific antigens of Blomia tropicalis, Tyrophagus putrescentiae, Acarus siro, and cockroach species Blattella germanica and Periplaneta americana were measured by ELISA.
Results HDM allergens were found in dust samples from bedding in 95% of the Chinese households. The median levels varied from <0.006 to 9.2 μg/g of dust, depending on the city. The percentages of households having HDM allergen levels associated with the risk of developing allergy sensitization and asthma were 65% and 25%, respectively. Specific antigens of the storage mite and cockroach were only found in samples from the southern and tropical regions of China. Levels of mite allergens were generally higher in samples from bedding compared to samples from the living room, even for storage mites, whereas levels of cockroach antigens were higher in the living room samples.
Conclusion HDM allergens are present in bedding dust samples from most Chinese households. Cities in southern and central China have relatively high levels of HDM major allergens compared to cities in northern and western China. Antigens of storage mites and cockroaches are not as common as HDM allergens.     

Comparison and Evaluation of Several Dermatophagoides Pteronyssinus Allergen Extracts for Skin Prick Test

Objective To evaluate the significance of several Dermatophagoides pteronyssinus allergen extracts for skin prick test （SPT） in patients allergic to Dermatophagoides pteronyssinus. Methods Two hundred and nineteen patients enrolled in Peking Union Medical College Hospital underwent SPT and serum specific IgE assay to detect the Dermatophagoides pteronyssinus allergen. Three kinds of house dust mite allergen extracts were used for SPT, including the Dermatophagoides pteronyssinus extract prepared by our laboratory （group A）, standardized Dermatophagoides pteronyssinus extract （group B）, and mixed extracts of Dermatophagoides pteronyssinus and Dermatophagoidesfarinae （group C）. Human serum specific IgE result was regarded as the reference standard for diagnosis of Dermatophagoides pteronyssinus allergy. The receiver operating characteristic （ROC） curve was used to evaluate the diagnostic performance of SPT with the extracts of three groups.Results SPT results showed that the median wheal diameter of group A, group B, and group C was 0.43, 0.35, and 0.28 cm, respectively, with significant difference among three groups （P〈0.05）. The difference was significant between group A and B （P〈0.01） as well as group A and C （P〈0.01）, but not between group B and C （P〉0.05）. There was no local urticaria or systemic allergic reactions following the procedure of SPT. Local reaction was observed in 5 patients and delayed reaction was in 2 patients of group A. As for group B and C, local reaction occurred in 3 cases and delayed reaction in 2 cases in each group. The area under ROC curve of SPT with extract in group A, group B, and group C was 0.765, 0.801, and 0.782, respectively. Based on the detection results of serum specific IgE, the sensitivity of SPT in diagnosis of Dermatophagoides pteronyssinus allergy with extract of group A, group B, and group C was 92.4%, 87.0%, and 81.5%, and the specificity was 60.6%, 73.2%, and 74.8%, respectively. Conclusion The Dermatophagoides pteronyssinus extract for SPT prepared by our laboratory offers good sensitivity and specificity comparable to commercially available allergen extracts, and it may be an appropriate candidate for clinical screening and diagnosis of Dermatophagoides pteronyssinus allergy.     

Prevalence of sensitivity to cockroach allergens and IgE cross- reactivity between cockroach and house dust mite allergens in Chinese patients with allergic rhinitis and asthma

Background Cockroaches are an important indoor allergen source causing allergic rhinitis and asthma. The aim of this study was to investigate the cockroach prevalence in mainland of China and the cross-reactivity of IgE between cockroach and house dust mite allergen in Chinese patients.Methods The cockroach sensitization pattern was based on a skin prick test （SPT） obtained from a national multicenter prevalence study, in which 6304 patients from 25 allergy centers across China participated. Factors, including different regions of China, age, gender and the correlations between the American and German cockroaches and house dust mite Der p were investigated. Eighteen out of 1236 clinical sera from south China were selected to perform the cross-inhibition assay between house dust mites and cockroaches.Results Totally 25.7% of patients were SPT positive to the American cockroach （Periplaneta Americana, Per a） and 18.7% SPT positive to the German cockroach （Blattella germanica, Bla g）. The prevalence of positive cockroach SPT was higher in southern than in northern China, higher in adults than in children, and higher in males than in females.Patients had relatively low levels of cockroach SPT reactions, mainly class 1 or 2. Of the SPT positive cockroach patients,88% were also SPT positive to house dust mite （Dermatophagoides pteronyssinus, Der p）. An IgE cross-inhibition study confirmed that Der P sensitization could cause false positive SPT reactions against cockroach.Conclusions A relatively high prevalence of cockroach sensitivity was found in mainland of China. However, a cross-inhibition study showed that only a small number of patients appear to have Bla g and/or Per a as primary sensitizing source. The importance of cockroaches as a risk factor for sensitization and triggers of allergic symptoms in mainland of China needs to be further investigated.     

尘螨变应原皮内试验与血清IgE检测结果的相关性

目的 探讨变应性鼻炎粉尘螨及户尘螨变应原体内检测与体外检测的相关关系.方法 选择变应性鼻炎患者34例, 体内检测采用皮内试验法, 体外检测采用血清特异性IgE检测法, 根据皮内试验结果分为:极强阳性组、强阳性组、中阳性组、阳性组及阴性组5个级别;应用Excel软件进行统计分析.结果 粉尘螨、户尘螨变应原皮内试验分别与体外血清lgE检测具有高度相关性;变应原皮内试验 (即户尘螨与粉尘螨的平均值) 与IgE检测具有高度相关性, 相关系数r=0.8532 (P<0.01, n=20) .结论 粉尘螨和户尘螨变应原皮内试验与体外血清lgE检测具有高度相关性, 临床上可以在患者条件允许的情况下先做皮内试验作为筛查, 然后再用体外检测进一步验证;对于高敏患者或其他不接受皮内试验的患者, 可直接进行体外检测.     

18岁以下年龄组过敏性鼻炎-哮喘综合征患者变应原检测结果调查分析

目的探讨低年龄组过敏性鼻炎-哮喘综合征患者的变应原特点。方法采用德国Allergy Screen Test Kit检测系统对515例18岁以下过敏性鼻炎-哮喘综合征患者进行血清总Ig E和19组变应原特异性Ig E定量检测,统计血清总Ig E的增高比例和变应原阳性率,分析其变应原特点。结果 515例18岁以下鼻炎-哮喘综合征患者血清总Ig E的增高比例为77.9%。本组变应原s Ig E阳性率前10位的变应原依次是:户尘螨(70.1%)、屋尘(34.0%)、霉菌(33.2%)、牛奶(24.1%)、狗毛(23.5%)、鸡蛋白(12.6%)、腰果(11.7%)、猫毛(10.9%)、早晚春花粉(8.0%)、夏秋花粉(7.4%)。不同年龄组变应原阳性率比较提示学龄前组(3~6岁)最高,达69.5%。结论本组中77.9%过敏性鼻炎-哮喘综合征患者具有过敏性体质。18岁以下过敏性鼻炎-哮喘综合征患者吸入性变应原阳性率前3位是户尘螨、屋尘、霉菌;食入性变应原阳性率前3位是牛奶、鸡蛋白、腰果。     

260例变态反应性皮肤病患者血清过敏原的检测和分析

目的 对变态反应性皮肤病患者血清进行过敏原检测及分析,以了解过敏原的种类.方法 采用免疫印迹法对260例变态反应性皮肤病患者血清总IgE及变应原特异性IgE水平进行检测,检测分为吸入组和食物组.结果 61.1%患者血清总IgE水平增高,66.9%患者血清特异性IgE增高.变应原特异性IgE吸入组以户尘螨阳性率最高,屋尘其次;食物组以蟹为最高,其后是牛奶.变应原特异性IgE阳性率高于总IgE阳性率.结论 过敏原与变态反应性皮肤病有一定相关性,因此过敏原的检测对其诊疗有重要价值.     

海口市儿童哮喘变应原流行病学调查研究

目的 了解海口市儿童哮喘常见变应原状况.方法 2012年9~12月本研究采取整群抽样的流行病学方法调查海口市0~14岁儿童10 241名,确诊哮喘患儿324例,对所有哮喘患儿进行过敏史问卷调查及皮肤点刺试验和血清过敏原特异性IgE (SIgE)抗体检测.结果 324例哮喘患儿中224例(69.1%)过敏史阳性,其中湿疹91例(28.1%),食物过敏64例(19.8%),药物过敏28例(8.6%),香烟过敏18例(5.6%),装修气味过敏19例(5.9%),宠物过敏13例(4.0%),油烟过敏11例(3.4%),花粉过敏9例(2.8%).皮肤点刺阳性率为83.0%(269例),其中屋尘螨及粉尘螨阳性率分别为90.1%(292例)及89.2%(289例),热带螨为85.5%(277例),蟑螂19.6%(64例),霉菌3.4%(11例),花粉3.4%(11例),猫毛11.7%(38例),狗毛25.3%(82例).吸入性过敏原IgE阳性率为72.2%(235例),以尘螨为最高(69.8%,226例);食入性过敏原IgE阳性率为28.7%(93例),牛奶占25.3%(82例),鸡蛋白占9.9%(32例),虾蟹占22.8%(74例),热带水果占9.0%(24例),鱼占8.3%(27例).不同年龄组哮喘患儿吸入性过敏原特异性IgE阳性率比较显示,3~7岁组屋尘螨、蟑螂阳性率高于0~3岁组(P<0.05),7~14岁组屋尘螨、蟑螂、狗毛上皮、猫毛上皮阳性率高于0~3岁组(P<0.05).结论 海口市儿童哮喘常见变应原以屋尘螨、粉尘螨和热带螨为主,狗毛、蟑螂及猫毛比例也较高.食入性过敏原以牛奶、鸡蛋白多见,且虾蟹、热带水果致敏对本地区儿童哮喘的影响也不容忽视.     

深圳特区变应性鼻炎患者螨与蟑螂过敏原体外检测与分析

目的探讨深圳特区变应性鼻炎患者血清中螨和蟑螂过敏原特异性免疫球蛋白E(sIgE)的情况及其相互关系。方法采用酶联免疫吸附法(ELISA)对该院1998~2010年就诊于耳鼻咽喉科门诊的1 935例患者进行血清过敏原体外检测。结果在变应性鼻炎患者中,对尘螨过敏的具有极高的阳性率,蟑螂过敏也有较强的阳性率,血清中的sIgE值尘螨显著高于蟑螂(P<0.01)。结论尘螨和蟑螂均为引起变应性鼻炎最主要的过敏原之一,尤其是尘螨,预防和治疗研究工作的重点应当是这些过敏原。     

武汉地区成人哮喘的过敏原检测及临床意义

目的了解武汉地区成人哮喘的过敏原。方法应用北京新华联协和药业有限责任公司提供的29种过敏原,对57例哮喘进行皮肤点刺试验。结果57例哮喘中可检出过敏原者50例,占87．7％,强阳性30例,占52．6％。武汉地区最常见5种过敏原是粉尘螨[41（71．9％）]、屋尘螨[38（66．7％）]、室内尘土[32（56．1％）]、蟑螂[25（43．9％）]和梧桐花粉[19（33．3％）]。结论武汉地区成人哮喘的发作与过敏原关系密切,尘螨是最常见的过敏原,梧桐花粉是最常见的致敏花粉。     

尘螨Ⅱ类改组变应原对哮喘小鼠免疫治疗的效果

目的 探讨以改组尘螨Ⅱ类变应原基因Der f2和Der p2后表达的蛋白为变应原,对小鼠哮喘模型的特异性免疫治疗效果。方法 随机将160只清洁级BALB/c小鼠分为PBS组（阴性对照组）,Der f2和Der p2免疫治疗组（阳性对照组）,哮喘组,改组变应原免疫治疗M01、M03、M08、M10蛋白治疗组。用尘螨提取液于0、7、14d腹腔注射致敏激发BALB/c小鼠,第21天雾化吸入激发,连续7d,其中各免疫治疗组于第25～27天雾化前30min分别用Der f2、Der p2和改组变应原进行特异性免疫治疗,PBS组则用PBS进行腹腔注射和雾化吸入。最后1次雾化24h后,引颈处死。分别观察肺组织病理变化、支气管肺泡灌洗液（BLAF）白细胞计数,酶联免疫吸附试验（ELISA）测定BLAF和脾细胞培养上清液细胞因子IL-4、IL-17和IFN γ的含量及血清中特异性IgG1、IgE抗体水平变化。结果 与哮喘组比较,改组变应原免疫治疗组和阳性治疗组肺部炎症显著减轻, BALF中的总细胞数及嗜酸性粒细胞数,血清中抗原特异性IgG1、IgE抗体均显著降低;免疫治疗组（包括阳性对照）的BALF和脾细胞培养上清液中的细胞因子IL-4、IL-17均明显降低,IFN-γ含量显著升高。结论 通过基因改组获得的尘螨Ⅱ类变应原改组疫苗免疫治疗小鼠哮喘,可有效降低尘螨引起的小鼠肺部炎症。     

1120例变态反应性皮肤病患者血清特异性IgE检测

目的探讨本地区变态反应性皮肤病常见过敏原。方法采用IVT过敏原体外测试系统对1120例变态反应性皮肤病患者的血清进行特异性IgE检测。结果吸入性血清过敏原总IgE阳性率63．39％,常见的吸入性过原依次为尘螨、粉螨、屋尘螨;点青霉菌,交链孢菌,黑根霉;食入性血清过敏原总IgE阳性率为68．75％,食入性血清过敏原为牛奶、鱼、蟹和虾,蛋白;大部分患者对两种以上的过敏原过敏。结论IVT可用于变态反应性皮肤病病因的检测,对临床诊断和治疗有重要的参考价值。     

广州市海珠区变态反应体质儿童过敏原调查

目的:获得广州市海珠区儿童的各类过敏原的分布特点,为本区儿童的"免疫照护"提供参考价值的资料。方法:将14岁以下有典型病史的特异性皮炎、荨麻疹、儿童慢性咳嗽、小儿哮喘、过敏性鼻炎等变态反应体质的儿童分成婴幼儿组和少儿组两组,利用荧光酶免法检测45种过敏原。结果:共获得371例阳性过敏原结果,其中户尘螨过敏原阳性128例,占34.5%,粉尘螨过敏原阳性95例,占25.6%,牛奶过敏原阳性75例,占20.2%;鸡蛋白过敏原阳性51例,13.7%。其中婴幼儿组以食入性过敏原牛奶和鸡蛋白为主,少儿组以吸入性过敏原尘螨为主,两组比较,P均<0.05,存在明显差异。结论:广州市海珠区儿童过敏性疾病的主要过敏原是户尘螨、粉尘螨、牛奶和鸡蛋白。婴幼儿是以食入性过敏原牛奶和鸡蛋白致敏为主,少儿以吸入性致敏原尘螨致敏为主。     

标准化免疫治疗对哮喘儿童屋尘螨特异性IgG4和肺功能的影响

目的 探讨标准化屋尘螨变应原制剂免疫治疗儿童哮喘,对其肺功能、气道反应性及血清屋尘螨特异性IgG4、血清屋尘螨特异性IgE的影响.方法 选择31例轻-中度屋尘螨过敏性哮喘儿童,给予标准化屋尘螨变应原制剂治疗,疗程25周,检测治疗前后肺功能指标FEV1和PEF占预计值的百分比,PC20、血清屋尘螨特异性IgG4和屋尘螨特...     

化工区变应性鼻炎变应原的特异性分析

目的了解宁波镇海化工区变应性鼻炎（AR）变应原情况,为高污染地区AR防治提供科学依据。方法采用宁波美康医学检验中心变应原体外检测试剂盒,以酶免疫分析法对该地区506例AR患者进行变应原血清总IgE（tIgE）和特异性IgE（sIgE）检测,其中镇海化工区274例、城区232例,并对两区域变应原IgE进行特异性分析。结果两个区域AR患者的变应原sIgE差异主要体现在吸入性变应原方面。化工区AR患者的尘螨、屋尘、霉菌、树类和艾蒿sIgE均显著高于城区患者（P〈0.05）,其中尘螨和屋尘差异最大（P〈0.01）。在两区域中,患者青少年组在尘螨、带鱼、虾蟹、龙虾、食品I和肉类sIgE方面要显著多于成年人组（P〈0.05）,而成年人组在屋尘、树类、蟑螂、艾蒿和皮屑sIgE方面要显著多于青少年组（P〈0.05）。结论受该地区气候潮湿及镇海化工区环境污染的影响,化工区AR变应原tIgE和sIgE较高,表明该地区AR较为严重,患病率可能较高。     

过敏性鼻炎血清特异性IgE与皮肤点刺试验的关联性分析

目的评估血清特异性IgE检测与皮肤点刺试验在诊断过敏性鼻炎中的关联性。方法皮肤点刺试验中尘螨阳性的过敏性鼻炎患者82例,采用瑞典法玛西亚公司UniCAP100系统检测血清尘螨特异性IgE,包括屋尘螨及粉尘螨,统计分析二者的关联性。结果82例尘螨过敏患者中,55例尘螨血清特异性IgE检测结果大于或等于1级,特异性IgE水平与皮肤点刺试验结果的等级呈正相关(屋尘螨rs=0.58,粉尘螨rs=0.60)。不同年龄段患者的血清特异性IgE检测结果与皮肤点刺试验的相关系数随年龄的增长而增大,40岁以上患者二者的检测结果无相关性。结论尘螨过敏性鼻炎患者血清特异性IgE水平与皮肤点刺试验结果呈正相关,且随患者年龄变化而有所差异。诊断40岁以上患者时,推荐同时进行2种检测。     

1491例变态反应性疾病患者过敏原检测结果分析

目的调查分析常州地区变态反应性疾病患者常见过敏原的分布情况,比较不同年龄阶段、不同季节的过敏原有无差异。方法选取本院2016年1月～2018年1月收治的1491例变态反应性皮肤病患者,采用16种过敏原对患者进行皮肤点刺试验,生理盐水作阴性对照,以组胺作阳性对照。结果 1491例变态反应性皮肤病患者常见过敏原阳性率从高到低依次为:粉尘螨89.67%、户尘螨86.38%、油菜花7.11%、葎草花粉6.37%蟑螂4.76%、猫毛3.76%。过敏原的季节性来分析:粉尘螨、户尘螨、夏季最高;春天比其他三个季节低;各种花粉在夏季最低,蟑螂夏季最高。粉尘螨、户尘螨、猫毛、狗上皮、狗毛季节性变化不是太明显。粉尘螨、户尘螨、猫毛、牛奶、鸡蛋黄未成年组高于成年组(P0.05)。葎草花粉、油菜花粉、蟑螂成年组高于未成年组(P0.05)。过敏原在同年龄阶段的男女中间的差异不明显(P0.05)。结论在常州地区,最主要的过敏原为粉尘螨、户尘螨、油菜花,过敏原在不同季节、不同年龄阶段分布不同,为本地区常见变态反应性疾病的防治提供科学依据,皮肤点刺试验用于明确变态反应性疾病致敏原因安全有效。     

小儿变应性鼻炎粉尘螨检测与脱敏治疗

目的观察与探讨小儿变应性鼻炎粉尘螨检测与脱敏治疗。方法本文选择了112例变应性鼻炎患儿,依照变应原的种类将其分成观察组70例与对照组42例。观察组仅对粉尘螨或是屋尘螨有过敏反应,对照组对尘螨及其他变应原均有过敏反应：所有患儿均通过标准化粉尘螨滴剂这种方式进行1年的舌下特异性免疫方法治疗（SLIT）并做好粉尘螨皮试检测。观察治疗前后两组患儿的症状评分与药物评分以及两组的治疗效果差异。结果所有患儿均完成治疗,治疗前后两组患儿在症状评分方面均得到了显著改善,在药物评分方面,两组患儿在治疗前后也得到了显著改善,两组比较差异有统计学意义（P〈0．05）,组间比较差异无统计学意义（P〉0．05）。结论将标准化粉尘螨滴剂放置舌下进行含服能够用于对尘螨合并有多种变应原合并症的变应性鼻炎的治疗,效果良好。     

HOUSE DUST MITE ALLERGEN （Derp1 AND Blot5） LEVELS IN ASTHMATICS＇ HOME IN HONGKONG

Objective To measure Derpl and Blot5 allergen levels in asthmatics' homes in Hongkong.Methods Seventy houses were enrolled for a mite indoor environment study. Dust samples were obtained from two sites of each patients' house: bed and floor. Derpl and Blot5 levels were quantified by a two-site monoclonal antibody-based ELISA technique.Results The levels of Derpl allergens found in bed (geometric mean (GM) 3.43 μg/g of dust; 95%CI, 1.89-4.96 μg/g)and on the floor (GM 1.12 μg/g of dust; 95%CI, 0.71-1.53 μg/g) indicated significant differences (P=0.005). However, the levels of Blot5 allergens found in bed (GM 19.00 μg/g of dust; 95%CI, 0.89-38.90 μg/g) and on the floor (GM 6.14 μg/g of dust; 95%CI, 0.40-11.90 μg/g) showed no statistically significant difference. In addition, in regards to the exposure index for Derpl and Blot5 allergens found in bed and on the floor, 17.6% in bed and 8.6% on the floor had levels of Blot5 ≥ 10 μg/g of dust, higher than those obtained for Derp1 (7.2% and 0% in bed and on the floor respectively, P＜ 0.05); higher percentages in bed and on the floor (25.0% and 35.7%) were observed for levels of Blot5 =0 μg/g of dust as compared with Derpl in bed and on the floor (4.3% and 14.5% respectively, P＜ 0.05).Conclusions Derpl and Blot5 are the major allergens found in this regional study, Blot5 is a more potent allergen in Hongkong, probably reflecting the high level of exposure to Blomia tropicalis (Bt). Bt and Dermatophagoides pteronyssinus (Dp) allergens should be included for precise diagnosis and effective immuno-therapeutic treatment of mite allergy in Hongkong.