摘要研究背景：随着生活方式和环境的改变，支气管哮喘（Bronchial asthma，BA，以下简称哮喘）的过敏原种类和其它触发因素增多，近年来，儿童哮喘的发病率快速上升。儿童哮喘发生时，体内各种细胞因子及相关蛋白相继发生一系列的动态变化，本研究尝试采用纳米磁珠结合SELDI-TOF-MS技术，对确诊为儿童哮喘和健康儿童的血清进行蛋白质组剖析，通过运算规则筛选能够区分儿童哮喘和健康者的血清蛋白差异峰，并初步构建儿童哮喘的诊断模型，为哮喘儿童的临床诊断提供新的方法，同时为相关差异蛋白峰的后续鉴定验证奠定基础。研究方法：利用弱阳离子(weak cation exchange, WCX)磁珠结合SELDI-TOF MS技术对76例哮喘儿童和84例健康儿童的血清样本进行分析获得血清蛋白指纹图谱，运用Biomarker Pattern Software (BPS)软件对数据进行筛选构建哮喘的诊断模型。研究结果：50例哮喘儿童和50例健康儿童血清蛋白指纹图谱的比较，显示40个蛋白峰在哮喘儿童和健康儿童之间存在差异 (P<0。05, Fold≥1。5)。筛选5349。7, 5821。9, 7845。6和33314。4 Da四个蛋白峰构建儿童哮喘诊断模型，其灵敏度为96。0％，特异度为84。0％。盲法检验的灵敏度为88。5％，特异度为82。4％。结论：利用纳米磁珠结合SELDI-TOF MS的哮喘蛋白指纹图谱检测技术筛选了与哮喘有关的差异蛋白峰，构建了哮喘儿童早期诊断的蛋白峰诊断模型，为哮喘儿童临床早期快速特异诊断的血清蛋白组学标准奠定初步基础。88665

Abstract Background: With the changing environment and lifestyle, the sensitization and triggering factors are increased which could induce bronchial asthma (BA, hereinafter referred to as asthma)。 In recent years, the incidence of asthma in children is increased rapidly。 When asthma occurs, a variety of cyto214kines and related proteins in children undergo a series of dynamic changes。 In this study, we tried to use proteomics analysis the serum from children with asthma and healthy volunteers by using SELDI-TOF MS combined with Weak cation exchange (WCX) magnetic beads。 And finding serum protein peaks to discriminate patients with asthma and healthy control subjects by calculation rules, and to establish new diagnosis models for the clinical characterization of asthma, and to provide foundation for the following identification of differential protein peaks。

Methods: SELDI-TOF MS combined with weak cation exchange (WCX) magnetic beads was used to screen serum samples from the 76 asthma and 84 healthy control subjects。 A classification model was established by Biomarker Pattern Software (BPS)。

Results: The protein profile of the serum samples from the 50 asthma and 50 healthy control subjects were analyzed with Biomarker Wizard software, 40 protein peaks were found to discriminate patients with asthma and healthy control subjects (P<0。05, Fold≥1。5), The diagnosis model based on the four peaks (5349。7, 5821。9, 7845。6 and 33314。4 Da) was established which could distinguish asthma from the healthy controls with the sensitivity of 96。0％ and the specificity of 84。0%。 And the blinding test could distinguish asthma from the healthy controls with the sensitivity of 88。5％ and the specificity of 82。4%。

Conclusions: SELDI-TOF MS combined with WCX magnetic beads can detect discriminating protein peaks and develop a diagnostic model for asthma with high sensitivity and specificity。 This provided a biological basis for rapid diagnosis of asthma using serum proteomics technology。

毕业论文关键词：哮喘；生物标记物；蛋白组学；表面加强激光解析电离飞行时间质谱

Key words: Asthma; Biomarker; Proteomics; SELDI-TOF MS

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