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Exploring the hepatitis C virus genome using single molecule real-time sequencing
- Source :
- World Journal of Gastroenterology
- Publication Year :
- 2019
- Publisher :
- Baishideng Publishing Group Inc., 2019.
-
Abstract
- Single molecular real-time (SMRT) sequencing, also called third-generation sequencing, is a novel sequencing technique capable of generating extremely long contiguous sequence reads. While conventional short-read sequencing cannot evaluate the linkage of nucleotide substitutions distant from one another, SMRT sequencing can directly demonstrate linkage of nucleotide changes over a span of more than 20 kbp, and thus can be applied to directly examine the haplotypes of viruses or bacteria whose genome structures are changing in real time. In addition, an error correction method (circular consensus sequencing) has been established and repeated sequencing of a single-molecule DNA template can result in extremely high accuracy. The advantages of long read sequencing enable accurate determination of the haplotypes of individual viral clones. SMRT sequencing has been applied in various studies of viral genomes including determination of the full-length contiguous genome sequence of hepatitis C virus (HCV), targeted deep sequencing of the HCV NS5A gene, and assessment of heterogeneity among viral populations. Recently, the emergence of multi-drug resistant HCV viruses has become a significant clinical issue and has been also demonstrated using SMRT sequencing. In this review, we introduce the novel third-generation PacBio RSII/Sequel systems, compare them with conventional next-generation sequencers, and summarize previous studies in which SMRT sequencing technology has been applied for HCV genome analysis. We also refer to another long-read sequencing platform, nanopore sequencing technology, and discuss the advantages, limitations and future perspectives in using these third-generation sequencers for HCV genome analysis.
- Subjects :
- Genome, Viral
Hepacivirus
Computational biology
Biology
Antiviral Agents
Genome
Deep sequencing
03 medical and health sciences
0302 clinical medicine
Drug Resistance, Multiple, Viral
Nanopore sequencer
Humans
Resistance-associated substitution
Gene
Sequence (medicine)
Whole genome sequencing
Hepatitis C virus
Haplotype
Gastroenterology
High-Throughput Nucleotide Sequencing
Minireviews
PacBio RSII
General Medicine
Hepatitis C
Single molecule real-time sequencing
030220 oncology & carcinogenesis
RNA, Viral
030211 gastroenterology & hepatology
Nanopore sequencing
Third generation sequencing
Single molecule real time sequencing
Subjects
Details
- ISSN :
- 10079327
- Volume :
- 25
- Database :
- OpenAIRE
- Journal :
- World Journal of Gastroenterology
- Accession number :
- edsair.doi.dedup.....38dd88f2910c949058c6442db57df479