【摘要】 目的:探索蛋白芯片技术检测乙型肝炎病毒(HBV)抗HBc-IgM的可行性。方法:应用Nano-Plotter TM-压电式微量喷墨点阵制备系统自制的核心抗原蛋白芯片在经10%山羊血清封闭后加入待测血清37℃孵育,PBST清洗晾干后后再次加入检测抗体即HRP-抗人Ig M抗体,芯片用伯乐成像仪检测有无信号。结果:24份HBs Ag和抗HBc阳性血清中经蛋白芯片检测抗HBc-IgM阳性率为83.3%(20/24);24份健康志愿者血清中经蛋白芯片检测抗HBc-IgM阳性率为4.1%(1/24)。乙肝血清与健康志愿者血清化学发光信号差异明显。结论:蛋白芯片技术可较好地应用于定性检测HBV抗HBc-IgM,为临床快速判断是否存在HBV感染和监测慢性肝炎HBV活动性提供新的辅助诊断方法。更多还原

【Abstract】 Objective: To explore the feasibility of protein microarray for detecting human anti-hepatitis B virus core antibody Ig M qualitatively. Methods: Protein microarray was fabricated by GeSiM Nano-PlotterTM Micropipetting System. The core antigen protein spotted on the slides with BSA spotted as control protein. All slides were blocked in a coupling buffer(10% normal goat serum with 0.1%NaN3). Then serum samples were added on the slides and incubated for 2 h at 37℃. After rinsed by PBST, the slides were added with HRP-anti-human Ig M antibody and then scanned by a chemiluminescent scanner, Chemi DocTM MP System(Bio-Rad,California, USA). Results: The positive rate of anti-HBc Ig M was 83.3%(20/24) in 24 HBs Ag and anti-HBc positive serum samples detected by protein microarray, while the positive rate of anti-HBc Ig M was 4.1%(1/24) in 24 healthy volunteers. The chemiluminescent signal difference was significant between hepatitis B positive and healthy serum samples. Conclusions: Protein microarray technology could be applied to detect anti-HBc Ig M qualitatively, and provide a new auxiliary diagnostic method for clinical rapid determination of HBV infection and monitoring HBV chronic hepatitis activity.更多还原