基于微阵列的病原体芯片，用于同时分子检测输血传播的传染性病原体。
A microarray-based pathogen chip for simultaneous molecular detection of transfusion-transmitted infectious agents.
机构信息
Infectious Diseases Section, Department of Transfusion Medicine, National Institutes of Health Clinical Center, Building 10, Room 1C711, 10 Center Drive, Bethesda, MD, 20892, USA.
出版信息
J Transl Med. 2019 May 14;17(1):156. doi: 10.1186/s12967-019-1905-4.
BACKGROUND
New and emerging transfusion-transmitted infections remain a threat to the blood supply. Blood donors are currently screened for less than half of known agents, primarily by individual tests. A screening platform that could simultaneously detect all known transfusion-transmitted pathogens and allow rapid addition of new targets would significantly increase blood safety and could improve the response to new agents. We describe the early stage development and validation of a microarray-based platform (pathogen chip) for simultaneous molecular detection of transfusion-transmitted RNA viruses.
METHODS
Sixteen RNA viruses that pose a significant risk for transfusion-transmission were selected for inclusion on the pathogen chip. Viruses were targeted for detection by 1769 oligonucleotide probes selected by Agilent eArray software. Differentially concentrated positive plasma samples were used to evaluate performance and limits of detection in the context of individual pathogens or combinations to simulate coinfection. RNA-viruses detection and concentration were validated by RT-qPCR.
RESULTS
Hepatitis A, B and C, Chikungunya, dengue 1-4, HIV 1-2, HTLV I-II, West Nile and Zika viruses were all correctly identified by the pathogen chip within the range of 10 to 10 copies/mL; hepatitis E virus from 10 to 10. In mixtures of 3-8 different viruses, all were correctly identified between 10 and 10 copies/mL.
CONCLUSIONS
This microarray-based multi-pathogen screening platform accurately and reproducibly detected individual and mixed RNA viruses in one test from single samples with limits of detection as low as 10 copies mL.
背景
新出现和新兴的经输血传播感染仍然是血液供应的威胁。目前，献血者的筛查范围不到已知病原体的一半，主要通过单项检测。如果有一种筛查平台能够同时检测所有已知的经输血传播病原体，并能快速添加新的检测目标，这将大大提高血液安全性，并能改善对新病原体的应对能力。我们描述了一种基于微阵列的平台（病原体芯片）的早期开发和验证，该平台用于同时分子检测经输血传播的 RNA 病毒。
方法
选择了 16 种可能经输血传播的 RNA 病毒，将其纳入病原体芯片。通过 Agilent eArray 软件选择的 1769 个寡核苷酸探针来针对病毒进行检测。使用差异浓缩的阳性血浆样本来评估单个病原体或组合模拟合并感染的情况下的性能和检测极限。通过 RT-qPCR 对 RNA 病毒的检测和浓度进行验证。
结果
甲型、乙型和丙型肝炎病毒、基孔肯雅热病毒、登革热病毒 1-4 型、HIV 1-2 型、HTLV I-II 型、西尼罗河病毒和寨卡病毒均在 10 至 10 拷贝/ml 的范围内通过病原体芯片正确识别；10 至 10 拷贝/ml 范围内可正确识别戊型肝炎病毒。在 3-8 种不同病毒的混合物中，所有病毒在 10 至 10 拷贝/ml 的范围内均能正确识别。
结论
这种基于微阵列的多病原体筛选平台能够从单个样本中一次测试准确、可重复地检测单个和混合的 RNA 病毒，检测极限低至 10 拷贝/ml。
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