应用表面增强激光解吸电离飞行时间质谱筛选急性特发性血小板减少性紫癜患儿血清生物标志物

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目的应用表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)技术结合蛋白芯片检测并筛选急性血小板减少性紫癜(AITP)早期诊断、病情变化监测及判断预后的血清蛋白质标志物,建立AITP诊断模型,并验证在血清蛋白质组水平,创立AITP的分子诊断新方法。方法应用SELDI-TOF-MS检测结合在蛋白质芯片上的血清蛋白质,获得35例AITP、35例健康对照者血清蛋白表达指纹图谱,并用Biomarker Wizard软件对数据进行分析,建立人工神经网络诊断模型。结果在分子质量/电荷比值(质荷比,M/E)为2 000~20 000,AITP组与健康对照组之间差异有统计学意义(P<0.01)的蛋白质峰有29个,其中高表达的蛋白质峰7个,低表达的蛋白质峰22个,从分析的结果看最有意义的蛋白质为质荷比为4 109.23、5 521.35的蛋白质,可以将其作为AITP的血清生物标志物;在蛋白质库中搜索与上述M/E值相对应的蛋白质,M/E为4 109.23的标志物是核蛋白体large subunit,M/E为5 521.35的标志物在蛋白质库中未发现。应用其血清标志物进入人工神经网络组合的诊断模型,将AITP组与健康儿童准确分组,敏感度为95%,特异度为90%。结论 1.M/E比为4 109.23、5 521.35的蛋白质组合的诊断模型能将AITP与健康儿童完全准确地分开,提示该蛋白质可能是AITP的血清生物标志物;其中M/E为5 521.35的蛋白质可能是新的蛋白质。2.SELDI-TOF-MS技术是寻找疾病相关性蛋白质的有效工具。 OBJECTIVE: To detect and screen the serum protein markers of early diagnosis of acute thrombocytopenic purpura (AITP), monitoring of disease changes and prognosis by SELDI-TOF-MS and protein chip to establish AITP diagnosis Model, and validate a new molecular diagnostic approach to AITP at the serum proteome level. Methods SELDI-TOF-MS was used to detect serum proteins bound to protein chips. Serum protein expression profiles of 35 AITP and 35 healthy controls were obtained. The data were analyzed by Biomarker Wizard to establish a diagnostic model of artificial neural network. Results There were 29 protein peaks between the AITP group and the healthy control group (P <0.01) with the mass / charge ratio (M / M) ranging from 2 000 to 20 000, of which 29 Seven protein peaks were expressed and 22 protein peaks were low expressed. From the results of the analysis, the most significant proteins were proteins with a mass-to-charge ratio of 4 109.23 and 5 521.35, which could be used as serum biomarkers of AITP. The protein library was searched for proteins corresponding to the above M / E values ​​with a M / E of 4 109.23 and a large subunit of nucleoprotein with a marker of M / E of 5221.35 was not found in the protein library. Using its serum markers into the diagnostic model of artificial neural network combination, the AITP group and healthy children were accurately grouped with a sensitivity of 95% and a specificity of 90%. Conclusion 1. The diagnostic model of protein combination with M / E ratios of 4 109.23 and 5 521.35 can completely and accurately separate AITP from healthy children, suggesting that the protein may be a serum biomarker of AITP; the M / E is 5221.35 Proteins may be new proteins. 2.SELDI-TOF-MS technology is an effective tool to find disease-related proteins.
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