Gene expression profile of peripheral blood mononuclear cells from mild and severe COVID-19 patients, severely ill COVID-19 patients treated with dexamethasone and healthy controls. Gene expression profile of peripheral blood mononuclear cells from mild and severe COVID-19 patients, severely ill COVID-19 patients treated with dexamethasone and healthy controls

The objective of this experiment was to compare the transcriptomic profile (NanoString platform) of peripheral blood mononuclear cells (PBMCs) from COVID-19 patients with mild disease, and patients with severe COVID-19 with and without dexamethasone treatment, and healthy controls. We analyzed PBMCs from 4 mild COVID patients, 3 severe COVID patients,4 severe COVID patients treated with dexamethasone, and 5 healthy controls Overall design: Total RNA was isolated from the patient and control PBMCs using the E.Z.N.A. total RNA isolation Kit (Omega Bio-tek).We analyzed 100 ng of RNA from four mild, three severe, and four severe COVID-19 patients treated with dexamethasone, along with five unexposed healthy donors using the human Host Response CodeSet panel (NanoString), which includes 773 genes covering the host immune response to infectious diseases, and 12 internal reference genes for data normalization. The RNA samples were hybridized as per the manufacturer’s instructions, and each sample was read in a multiplexed reaction with the reference (positive, negative controls, and housekeeping genes) probes on the NanoString nCounter Flex instrument.

本实验旨在比较轻症新型冠状病毒肺炎（COVID-19）患者、伴或不伴地塞米松治疗的重症COVID-19患者，以及健康对照者的外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）的转录组谱，检测基于NanoString平台开展。本研究共纳入4例轻症COVID-19患者、3例重症COVID-19患者、4例接受地塞米松治疗的重症COVID-19患者，以及5例健康对照者的PBMCs用于后续分析。整体实验设计如下：使用E.Z.N.A.总RNA提取试剂盒（Omega Bio-tek）从受试者及健康对照的PBMCs中提取总RNA。随后，取上述4例轻症、3例重症、4例接受地塞米松治疗的重症COVID-19患者，以及5例未暴露的健康供者的100 ng RNA，采用NanoString公司的人类宿主应答编码集芯片（human Host Response CodeSet panel）进行检测；该芯片包含773个覆盖宿主对传染病免疫应答的基因，以及12个用于数据标准化的内参基因。按照试剂盒制造商的操作说明完成RNA样本杂交，随后在NanoString nCounter Flex仪器上，通过多重反应对每个样本与参照探针（阳性对照、阴性对照及持家基因探针）进行信号读取。