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J Clin Microbiol. 2019 Jun 12. pii: JCM.00119-19. doi: 10.1128/JCM.00119-19. [Epub ahead of print]

Characterization and Clinical Significance of Natural Variability in Hepatitis B Virus Reverse Transcriptase in Treatment-naïve Chinese Patients by Sanger Sequencing and Next-generation Sequencing.

Fu Y1,2, Zeng Y1,2, Chen T1,2, Chen H1,2, Lin N1,2, Lin J1,2, Liu X1,2, Huang E1,2, Wu S1,2, Wu S1,2, Xu S1,2, Wang L1,2, Ou Q3,2.

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First Clinical College, Fujian Medical University, Fuzhou, China.
Department of Laboratory Medicine, The First Affiliated Hospital of Fujian Medical University, Fuzhou, China.
First Clinical College, Fujian Medical University, Fuzhou, China


Background: Mutations in HBV RT are associated with NA resistance during the long-term antiviral treatment. However, the characterization of mutations in HBV RT in untreated patients has not yet been well illustrated. Objective: To investigate the characterization and clinical significance of natural variability in HBV RT in treatment-naïve patients. Methods: HBV RT sequences were analyzed in 427 patients by sanger sequencing and 66 patients by next-generation sequencing. Results: Primary or secondary NAr mutations were not found except rtA181T by sanger sequencing, but detected by next-generation sequencing. Mutations were found in 56 RT AA sites by sanger sequencing, in 36 of which mutations could lead to changes of B or T cell epitopes in RT or S protein. The distribution of mutations was diverse in different sections with the RT region. Multiple mutations showed signifcant association with HBV DNA, HBsAg, HBeAg, age and severity of liver fibrosis. Mutations at rt251, rt266, rt274, rt280, rt283, rt284 and rt286 were found most in ALD group by next-generation sequencing. Conclusion: The present study demonstrates that NGS was more suitable than sanger sequencing for monitor NAr mutations in a low rate in the treatment-naïve patients and mutations in RT region might be involved in the progression to ALD.

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