Nano Medical Engineering Laboratory, RIKEN, Wako, Saitama, Japan.
Nano Medical Engineering Laboratory, RIKEN, Wako, Saitama, Japan ; Consonal Biotechnologies Co., Ltd., Funabashi, Chiba, Japan.
PLoS One. 2013 Dec 18;8(12):e81726. doi: 10.1371/journal.pone.0081726. eCollection 2013.
We developed an automated diagnostic system for the detection of virus-specific immunoglobulin Gs (IgGs) that was based on a microarray platform. We compared efficacies of our automated system with conventional enzyme immunoassays (EIAs). Viruses were immobilized to microarrays using a radical cross-linking reaction that was induced by photo-irradiation. A new photoreactive polymer containing perfluorophenyl azide (PFPA) and poly(ethylene glycol) methacrylate was prepared and coated on plates. Inactivated measles, rubella, mumps, Varicella-Zoster and recombinant Epstein-Barr viruse antigen were added to coated plates, and irradiated with ultraviolet light to facilitate immobilization. Virus-specific IgGs in healthy human sera were assayed using these prepared microarrays and the results obtained compared with those from conventional EIAs. We observed high correlation (0.79-0.96) in the results between the automated microarray technique and EIAs. The microarray-based assay was more rapid, involved less reagents and sample, and was easier to conduct compared with conventional EIA techniques. The automated microarray system was further improved by introducing reagent storage reservoirs inside the chamber, thereby conserving the use of expensive reagents and antibodies. We considered the microarray format to be suitable for rapid and multiple serological diagnoses of viral diseases that could be developed further for clinical applications.
我们开发了一种基于微阵列平台的用于检测病毒特异性免疫球蛋白 G(IgG)的自动化诊断系统。我们比较了我们的自动化系统与传统酶免疫测定(EIA)的功效。通过光辐照诱导的自由基交联反应将病毒固定到微阵列上。制备了一种含有全氟苯基叠氮化物(PFPA)和聚乙二醇甲基丙烯酸酯的新型光反应性聚合物,并涂覆在板上。将灭活的麻疹、风疹、腮腺炎、水痘带状疱疹和重组 EBV 病毒抗原添加到涂覆的板中,并用紫外线照射以促进固定化。使用这些制备的微阵列检测来自健康人血清的病毒特异性 IgG,并将获得的结果与来自传统 EIA 的结果进行比较。我们观察到自动化微阵列技术与 EIA 之间的结果具有高度相关性(0.79-0.96)。与传统 EIA 技术相比,基于微阵列的测定法更快,所需试剂和样品更少,操作更简单。通过在腔室内引入试剂储存库,进一步改进了自动化微阵列系统,从而节省了昂贵试剂和抗体的使用。我们认为微阵列格式适合用于病毒病的快速和多种血清学诊断,可以进一步开发用于临床应用。