Division of Infectious Diseases, The Miriam Hospital, Providence, Rhode Island; Division of Infectious Diseases, Warren Alpert School of Medicine, Brown University, Providence, Rhode Island.
Division of Infectious Diseases, Warren Alpert School of Medicine, Brown University, Providence, Rhode Island.
J Mol Diagn. 2021 Jun;23(6):719-731. doi: 10.1016/j.jmoldx.2021.02.008. Epub 2021 Mar 9.
Viral infections are major causes of morbidity and mortality in solid-organ and hematopoietic stem cell transplant recipients. This study evaluated the performance of the Galileo Pathogen Solution metagenomics Next-Generation sequencing assay to detect and quantify 11 DNA viruses (cytomegalovirus, Epstein-Barr virus, BK virus, human adenovirus, JC virus, herpes simplex virus 1 and 2, varicella zoster virus, human herpesvirus 6A and 6B, and parvovirus B19) and to qualitatively detect torque teno virus. DNA extracted from 47 plasma samples of viremic transplant recipients were subjected to DNA library preparation with pathogen enrichment/human background depletion, sequencing, and automated data analysis. The viral loads were determined with the Galileo assay using a standard curve generated from a calibration panel. All of the samples tested had a 100% agreement with the real-time quantitative PCR (qPCR) assays in detecting the primary virus targets and the majority of the quantified samples had a viral load difference within 0.46 log IU/mL or copies/mL. The mean difference for cytomegalovirus between the Galileo and qPCR assays was 0.21 log IU/mL (SD, ±0.43 log IU/mL). The mean difference for BK virus between the Galileo and qPCR assays was 0.17 log cp/mL (SD, ±0.67 log cp/mL). Additionally, 75 co-infections were detected in 31 samples by the Galileo assay. The study findings show that the Galileo assay can simultaneously detect and quantify multiple viruses in transplant recipients with results that are comparable with standard-of-care qPCR assays.
病毒感染是实体器官和造血干细胞移植受者发病率和死亡率的主要原因。本研究评估了 Galileo 病原体解决方案宏基因组下一代测序检测和定量 11 种 DNA 病毒（巨细胞病毒、EB 病毒、BK 病毒、人腺病毒、JC 病毒、单纯疱疹病毒 1 和 2、水痘带状疱疹病毒、人疱疹病毒 6A 和 6B 以及细小病毒 B19）的性能，并定性检测 torquetenovirus。对 47 份病毒血症移植受者的血浆样本进行 DNA 提取，然后进行病原体富集/人类背景耗竭的 DNA 文库制备、测序和自动数据分析。使用从校准面板生成的标准曲线，使用 Galileo 检测确定病毒载量。所有测试样本与实时定量 PCR（qPCR）检测主要病毒靶标完全一致，大多数定量样本的病毒载量差异在 0.46 log IU/mL 或拷贝/mL 以内。Galileo 和 qPCR 检测之间的巨细胞病毒平均差异为 0.21 log IU/mL（SD，±0.43 log IU/mL）。Galileo 和 qPCR 检测之间的 BK 病毒平均差异为 0.17 log cp/mL（SD，±0.67 log cp/mL）。此外，Galileo 检测在 31 个样本中检测到 75 种共感染。研究结果表明，Galileo 检测可同时检测和定量移植受者中的多种病毒，其结果与标准护理 qPCR 检测相当。
