Evaluation of a Next-Generation Sequencing Metagenomics Assay to Detect and Quantify DNA Viruses in Plasma from Transplant Recipients.
2021
Abstract 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 (Galileo) to detect and quantify 11 DNA viruses (CMV, EBV, BKV, HAdV, JCV, HSV-1, HSV-2, VZV, HHV-6A, HHV-6B and parvovirus B19) and to qualitatively detect TTV. 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 the samples tested had a 100% agreement with the qPCR assays in detecting the primary virus targets and the majority of the quantified samples had a viral load difference within 0.46log10 IU/mL or copies/mL. The mean difference for CMV between the Galileo and qPCR assays was 0.21 log10IU/mL (SD±0.43log10 IU/mL). The mean difference for BKV between the Galileo and qPCR assays was 0.17 log10cp/mL (SD±0.67log10cp/mL). Additionally, 75 co-infections were detected in 31 samples by the Galileo assay. The study findings demonstrate that the Galileo assay can simultaneously detect and quantify multiple viruses in transplant recipients with results that are comparable to standard of care qPCR assays.
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