Division of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, UK.
Anal Bioanal Chem. 2011 Nov;401(8):2549-59. doi: 10.1007/s00216-011-5332-5. Epub 2011 Sep 1.
Hepatitis C virus (HCV) is a major cause of chronic liver disease and liver cancer, and remains a large health care burden to the world. In this study we developed a DNA microarray test to detect HCV RNA and a protein microarray to detect human anti-HCV antibodies on a single platform. A main focus of this study was to evaluate possibilities to reduce the assay time, as a short time-to-result (TTR) is a prerequisite for a point-of-care test. Significantly reducing hybridisation and washing times did not impair the assay performance. This was confirmed first using artificial targets and subsequently using clinical samples from an HCV seroconversion panel derived from a HCV-infected patient. We were able to reduce the time required for the detection of human anti-HCV antibodies to only 14 min, achieving nanomolar sensitivity. The protein microarray exhibited an analytical sensitivity comparable to that of commercial systems. Similar results were obtained with the DNA microarray using a universal probe which covered all different HCV genotypes. It was possible to reduce the assay time after PCR from 150 min to 16 min without any loss of sensitivity. Taken together, these results constitute a significant step forward in the design of rapid, microarray-based diagnostics for human infectious disease, and show that the protein microarray is currently the most favourable candidate to fill this role.
丙型肝炎病毒（HCV）是导致慢性肝病和肝癌的主要原因，也是全球范围内的一个重大医疗保健负担。在这项研究中，我们开发了一种 DNA 微阵列测试来检测 HCV RNA 和一种蛋白质微阵列来检测人类抗 HCV 抗体在单个平台上。这项研究的一个主要重点是评估缩短检测时间的可能性，因为即时检测需要较短的检测时间（TTR）。显著缩短杂交和洗涤时间并不会影响检测性能。这首先使用人工靶标进行了验证，然后使用来自 HCV 感染患者的 HCV 血清转换面板的临床样本进行了验证。我们能够将检测人类抗 HCV 抗体所需的时间缩短至仅 14 分钟，实现了纳摩尔级的灵敏度。蛋白质微阵列的分析灵敏度与商业系统相当。使用涵盖所有不同 HCV 基因型的通用探针进行 DNA 微阵列也得到了类似的结果。在不损失灵敏度的情况下，PCR 后的检测时间从 150 分钟缩短至 16 分钟。总的来说，这些结果在设计用于人类传染病的快速、基于微阵列的诊断方面迈出了重要的一步，并且表明蛋白质微阵列目前是最适合发挥这一作用的候选者。
