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目的建立激素抗体组合微阵列检测方法,为垂体腺瘤患者及无症状高风险人群提供血清学早期筛查的新方法。方法通过对微阵列芯片制备条件进行摸索,选择出泌乳素、生长激素和促甲状腺素的单克隆抗体作为芯片反应时的最佳制备条件及检测体系后,将其合理组合固定在固相载体的表面而制备出低密度激素抗体蛋白微阵列,建立激素抗体组合微阵列芯片检测法,并对该方法的特异性、灵敏度、重复性、精密度和交叉反应进行初步评价,比较其与AxSYM全自动免疫分析仪的相关性。结果成功确定了激素抗体组合微阵列的制作条件,包括选择琼脂糖凝胶玻片作为点样载体,确定3种激素抗体包被浓度、点样缓冲液、固定时间等;优化了反应体系中的各种条件;得出了多种激素抗体组合微阵列检测方法从制作到反应结果分析的完整体系。结论本研究初步建立了多种激素抗体组合微阵列检测方法,并对其进行了方法学评价,该方法学的建立不仅为临床激素检测提供了一种新途径,也为以后将更多的相关检测项目纳入研究创造了条件。
OBJECTIVE: To establish a method for the combination of hormone antibody and microarray to provide early serological screening for patients with pituitary adenoma and asymptomatic high-risk individuals. Methods The preparation conditions of microarray chips were explored and the monoclonal antibodies against prolactin, growth hormone and thyroid stimulating hormone were selected as the optimal preparation conditions and detection system for the chip reaction, and then they were rationally combined and immobilized on the solid phase carrier Density hormone antibody protein microarray was prepared to establish a hormone antibody combinatorial microarray chip detection method, and the specificity, sensitivity, repeatability, precision and cross-reactivity of the method were evaluated, compared with AxSYM fully automatic Immunoassay relevance. Results The conditions for the production of hormone antibody combinatorial microarray were successfully established, including the selection of agarose glass slide as spotting carrier, the determination of the concentration of three kinds of hormone antibody coating, spotting buffer, fixing time and so on. The optimal reaction conditions Various conditions; obtained a variety of hormone antibody combinatorial microarray detection method from the production to the analysis of the results of a complete system. Conclusion This study established a number of combinations of hormone antibody combinatorial microarray method, and its methodological evaluation, the establishment of this method not only for clinical hormone detection provides a new way, but also for the future will be more relevant Test items to create the conditions for the study.