Evaluation of PCR-RFLP in the Pre-S Region as Molecular Method for Hepatitis B Virus Genotyping

Ouneissa, Rim; Bahri, Olfa; Yahia, Ahlem Ben; Touzi, Henda; Azouz, MM; Mami, N. Ben; Triki, Henda

doi:10.5812/hepatmon.11781

Cited by 5 publications

(4 citation statements)

References 46 publications

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“…Genotyping showed that genotype D was detected in the 5 studied patients. This genotype was previously described as a predominant HBV genotype in Tunisia and the Maghreb region as well as in the Middle East with a low co-circulation rate of genotype E (Ayed et al, 2007;Ezzikouri et al, 2008;Ouneissa et al, 2013).…”

Section: Discussionmentioning

confidence: 90%

Whole genome analysis of hepatitis B virus before and during long-term therapy in chronic infected patients: Molecular characterization, impact on treatment and liver disease progression

et al. 2022

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Hepatitis B virus (HBV) infection remains a serious public health concern worldwide despite the availability of an efficient vaccine and the major improvements in antiviral treatments. The aim of the present study is to analyze the mutational profile of the HBV whole genome in ETV non-responder chronic HBV patients, in order to investigate antiviral drug resistance, immune escape, and liver disease progression to Liver Cirrhosis (LC) or Hepatocellular Carcinoma (HCC). Blood samples were collected from five chronic hepatitis B patients. For each patient, two plasma samples were collected, before and during the treatment. Whole genome sequencing was performed using Sanger technology. Phylogenetic analysis comparing the studied sequences with reference ones was used for genotyping. The mutational profile was analyzed by comparison with the reference sequence M32138. Genotyping showed that the studied strains belong to subgenotypes D1, D7, and D8. The mutational analysis showed high genetic variability. In the RT region of the polymerase gene, 28 amino acid (aa) mutations were detected. The most significant mutations were the pattern rtL180M + rtS202G + rtM204V, which confer treatment resistance. In the S gene, 35 mutations were detected namely sP120T, sT126S, sG130R, sY134F, sS193L, sI195M, and sL216stop were previously described to lead to vaccine, immunotherapy, and/or diagnosis escape. In the C gene, 34 mutations were found. In particular, cG1764A, cC1766G/T, cT1768A, and cC1773T in the BCP; cG1896A and cG1899A in the precore region and cT12S, cE64D, cA80T, and cP130Q in the core region were associated with disease progression to LC and/or HCC. Other mutations were associated with viral replication increase including cT1753V, cG1764A/T, cC1766G/T, cT1768A, and cC1788G in the BCP as well as cG1896A and cG1899A in the precore region. In the X gene, 30 aa substitutions were detected, of which substitutions xT36D, xP46S, xA47T, xI88F, xA102V, xI127T, xK130M, xV131I, and xF132Y were previously described to lead to LC and/or HCC disease progression. In conclusion, our results show high genetic variability in the long-term treatment of chronic HBV patients causing several effects. This could contribute to guiding national efforts to optimize relevant HBV treatment management in order to achieve the global hepatitis elimination goal by 2030.

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Section: Discussionmentioning

confidence: 90%

Whole genome analysis of hepatitis B virus before and during long-term therapy in chronic infected patients: Molecular characterization, impact on treatment and liver disease progression

et al. 2022

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“…It is important to take into account the genetic variability of HBV when identifying the polymorphisms that modify RFLP patterns present at each restriction enzyme recognition site. Nevertheless, RFLP continues to be used for HBV genotyping (32,35,36).…”

Section: Discussionmentioning

confidence: 99%

“…Various RFLP protocols for HBV genotyping have been described, most of them based on the amplification and restriction of ORF S sequences (23,(27)(28)(29). The RFLP method is used in genotyping studies because it is a simple and relatively inexpensive method to determine the HBV genotype, particularly for large-scale analyses (30)(31)(32).…”

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confidence: 99%

Analysis of hepatitis B virus genotypes by restriction fragment length polymorphism

et al. 2015

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“…In Eastern Europe and Mediterranean countries, the most prevalent is genotype D (6). According to literature reports, along with genotype C, genotype D produces a more aggressive course of the disease and brings lower response rates to antiviral therapy (7)(8)(9)(10). Literature focused on this issue explains that mutations in the S gene could lead to structural and functional alter-ations in the HBV reverse transcriptase, with potential influence on viral replication capacity and efficacy of antiviral drugs, the escape mutants eluding the immune system, and explaining the coexistence of HBsAg and anti -HBsAb (11,12).…”

Section: Introductionmentioning

confidence: 99%

Concomitant Serum Presence of Hepatitis B Surface Antigen (HBsAg) and High Titers of Hepatitis B Surface Antibodies (Anti - HBsAb) in a Patient with Chronic Hepatitis B (HBV) Genotype D from Black Sea Coast Region: A Case Report

et al. 2018

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Introduction: Hepatitis B Virus genotypes influence chronic hepatitis B evolution and show geographic preferences. In Eastern Europe, HBV-D genotype seems to be dominant, still, without enough information regarding the prevalence of its four subtypes. In addition, treatment with Nucleos(t)ide Analogues (NUCs) has lower impact on HBV-D genotype clearance compared to others. This study aimed at presenting the case of a middle age female diagnosed, followed-up and treated for eight years with entecavir 0.5 mg/day, in whom an increase of anti-HBsAb titer was noted over the immunogenicity level and undetectable viremia, despite the continuous presence of HBsAg. This is an uncommon type of evolution, most patients with anti-HBsAb over 10 IU/mL show seroconversion in the "s" system in a few months. Case Presentation: The researchers used the COBAS TaqMan HBV Monitor Test (Roche Diagnostics, Branchburg, NJ) in order to measure serum HBV DNA level, then, the viral DNA was extracted from 200 µL of serum using QIAamp DNA blood mini kit (Qiagen, Germany). The amplification and sequencing of full DNA length was done by the Rolling Circle Amplification (RCA) technique. Hepatitis B Virus genotype was determined using the NCBI genotyping tool and phylogenetic analysis. Conclusions: The patient was proved to have D genotype. The DNA analyzes showed escape mutations in the "a" determinant within the S gene, represented by sQ129R, respectively, sI134T, reported by the literature in a small number of C genotype patients. The paradoxical serum profile of the current patient with positive HBsAg, increased titers of anti-HBsAb, and undetectable viral load could be a possible model of evolution for D genotype HBV chronic infected patients treated with NUCs, cohort genetic studies on patients with the same serum profile are required to confirm this pattern of evolution.

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Evaluation of PCR-RFLP in the Pre-S Region as Molecular Method for Hepatitis B Virus Genotyping

Cited by 5 publications

References 46 publications

Whole genome analysis of hepatitis B virus before and during long-term therapy in chronic infected patients: Molecular characterization, impact on treatment and liver disease progression

Whole genome analysis of hepatitis B virus before and during long-term therapy in chronic infected patients: Molecular characterization, impact on treatment and liver disease progression

Analysis of hepatitis B virus genotypes by restriction fragment length polymorphism

Concomitant Serum Presence of Hepatitis B Surface Antigen (HBsAg) and High Titers of Hepatitis B Surface Antibodies (Anti - HBsAb) in a Patient with Chronic Hepatitis B (HBV) Genotype D from Black Sea Coast Region: A Case Report

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