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Interdisciplinary Collaboration to Develop a Custom Genomic Analysis Pipeline for the Clinical Laboratory: Hepatitis
Gordon Ritchie1,2, Mahdi Mobini1,3, Venkat S Malladi4
1Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.
Background:
Next-generation sequencing for hepatitis B virus (HBV) and cytomegalovirus (CMV) antiviral drug resistance (AVDR) testing improves the sensitivity of variant detection, but availability of bioinformatics and analytical pipelines are key barriers to implementation.
Methods:
Plasma was extracted on MagNA Pure 24 (Roche Diagnostics) and next-generation sequencing performed on GridION (Oxford Nanopore Technologies) with R.10.4.1 flowcells. An in-house bioinformatics pipeline was developed using Nextflow and deployed on Microsoft Azure to process FASTQ files and automate reporting of HBV genotype and AVDR, as well as CMV AVDR (UL97/54).
Results:
A total of 71 samples for HBV genotyping and AVDR testing and 56 samples for CMV AVDR testing were analyzed and compared to reference pipelines (DeepChek® HBV and CMV). All HBV genotypes and resistant mutations were concordant. For CMV, 74 mutations were identified in the UL97/54 region by both pipelines. However, our in-house developed method identified an additional UL97 drug resistant mutation (del598-603) in one sample.
Conclusions:
A custom bioinformatics pipeline was developed for HBV and CMV genotyping and AVDR sequencing, which could be adapted to other targets to enable our clinical laboratory to expand clinical testing using next-generation sequencing.
