Evaluation of sequencing of HCV core/E1, NS5A and NS5B as a genotype predictive tool in comparison with commercial assays targeting 5′UTR

McCormick, A.L., Macartney, M.J., Abdi-Abshir, I., Labbett, W., Smith, C., Irish, D., Webster, D.P. and Haque, T. 2015. Evaluation of sequencing of HCV core/E1, NS5A and NS5B as a genotype predictive tool in comparison with commercial assays targeting 5′UTR. Journal of Clinical Virology. 66 (May), pp. 56-59. doi:10.1016/j.jcv.2015.03.006

TitleEvaluation of sequencing of HCV core/E1, NS5A and NS5B as a genotype predictive tool in comparison with commercial assays targeting 5′UTR
AuthorsMcCormick, A.L., Macartney, M.J., Abdi-Abshir, I., Labbett, W., Smith, C., Irish, D., Webster, D.P. and Haque, T.
Abstract

Background

Hepatitis C virus (HCV) genotyping is required for tailoring the dose and duration of antiviral therapy, predicting virological response rates, and selecting future treatment options.
Objective

To establish whether baseline genotypes, performed by INNO-LiPA Version 1.0 (v1.0), before 2008, were valid for making treatment decisions now or whether genotypic determination should be repeated. Furthermore, to evaluate concordance between Abbott RealTime genotype II assay (RT) and genotyping by sequencing HCV C/E1, NS5A, NS5B.
Study design

Genotyping by RT and sequencing was performed on paired historic and current specimens from 50 patients previously baseline genotyped using INNO-LiPA.
Results

Of 100 samples from 50 patients, ≥2 of HCV genomic target regions yielded a sequence that was suitable for genotyping, with 100% concordance, providing no evidence of recombination events. Genotype and subtype prediction based on RT and sequencing agreed in 62.8% historic and 72.7% current specimens, with a kappa coefficient score of 0.48 and 0.76, respectively.

LiPA could not subtype 46% of HCV gt1 infections, and LiPA subgenotype was only in agreement with RT and sequencing in 28.6% cases, where matched baseline and historic specimens were available.

Three patients were indeterminate by RT, and five patients with HCV gt1 infections could not be subtyped by RT. However, RT revealed mixed infections in five patients where sequencing detected only single HCV infection at 20% threshold.
Conclusion

Genotyping by sequencing, exhibited excellent concordance, with moderate to good agreement with RT, and could resolve RT indeterminates and subtype HCV-gt1 infections not possible by LiPA.

Keywords Hepatitis C virus; Genotyping; Subtyping; RealTime PCR; DNA sequencing; LiPA
JournalJournal of Clinical Virology
Journal citation66 (May), pp. 56-59
ISSN1386-6532
Year2015
PublisherElsevier
Digital Object Identifier (DOI)doi:10.1016/j.jcv.2015.03.006
Web address (URL)http://www.sciencedirect.com/science/article/pii/S1386653215000839
Publication dates
PublishedMay 2015

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