抗可提取性核抗原抗体蛋白质微阵列芯片的临界参考值研究

The study of protein microarray chip criticality reference values of anti-extractability-nucleoantigen(ENA) antibody

目的建立自身抗体蛋白质微阵列芯片化学发光法的临界参考值研究方法。方法利用阳性对照品和"临界值公司一级参考品"的信号比值确定各自身抗体指标的"临界值系数",利用每次实验中阳性对照品的信号值与"临界值系数"确定各自身抗体指标的临界参考值(cut-off),并对实验结果作判定。结果利用此方法确定的抗可提取性核抗原(ENA)抗体蛋白质微阵列芯片的临界参考值,使产品中各指标的检测特异度均达到98.75%以上,产品在混合性结缔组织病、系统性红斑狼疮、干燥综合征、进行性系统硬化症、多肌炎/皮肌炎病例样本中阳性检出率分别达到了100%、100%、95.56%、100%、100%和100%。结论本研究建立的自身抗体蛋白质微阵列芯片化学发光法临界参考值确定方法是科学合理的,有效地消除了实验过程中人为因素,判定结果具有可靠性、可重复性。

Objective To establish a method for determining the criticality reference values of a protein chip system（protein microarray with chemiluminescence method）for the detection of auto-antibodies.Methods The signal ratios of the positive control sample and the first degree cutoff reference material from the manufacturer were used to obtain the ＂cutoff index＂ for each auto-antibody biomarker.The ＂cutoff index＂ was then used together with the actual signal in each experiment to determine the actual cutoff value for each auto-antibody biomarker.These cutoff values were then used to determine the positive and negative results for each biomarker in the experiment.Results When the clinical cutoff values established using above method were used in a protein microarray system for the detection of anti-extractable nuclear antigens（ENA） antibodies,specificities for each auto-antibody were all above 98.75%.The positive detection rates for mixed connective tissue diseases,systemic lupus erythematosus,Sjogren syndrome,progressive systemic sclerosis,polymyositis/dermatomyositis were 100%,100%,95.56%,100%,100% and 100% respectively.Conclusion This study has established a valid method for the determination of cutoff values for a protein microarray system for the detection of auto-antibodies.This method eliminates the influence of human factors during the experiment and thus helps achieve reliable and repeatable results.