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Updated: Aug 23, 2026
Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection
Published on: November 7, 2018
Detection of hepatitis B virus core mutants by PCR-RFLP in chronically infected patients
M F Barros1, P A Rodrigues, S R Matias
1Immunology Department, Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo de Santana 130, 1169-056 Lisbon, Portugal. imunologia@fcm.unl.pt
Insights
Chronic hepatitis B virus (HBV) infection is a significant health concern. This study identified common mutations in the HBV core antigen gene, offering a rapid detection method for viral mutants in patients.
Area of Science:
- Virology
- Immunology
- Molecular Biology
Background:
- Chronic hepatitis B virus (HBV) infection poses a major global health challenge.
- The HBV core antigen is vital for immune recognition, influencing infection outcomes.
- Understanding HBV genetic variations is crucial for managing chronic infection.
Purpose of the Study:
- To detect and characterize point mutations within the HBV core open reading frame (ORF).
- To evaluate the frequency of specific HBV core gene mutations in chronically infected individuals.
- To establish a rapid and reliable method for identifying HBV mutants.
Main Methods:
- Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) was employed to analyze HBV DNA from patient serum.
- Direct sequencing confirmed PCR-RFLP findings for HBV DNA from chronically infected patients.
- A longitudinal study design was used to track viral changes in a subset of patients.
Main Results:
- Frequent mutations were identified at positions 2248 and 2147 (designated A3), and at position 2038 (designated M2).
- The wild-type HBV core genotype was present in approximately 50% of the analyzed samples.
- PCR-RFLP proved to be a rapid and reliable technique for mutation detection.
Conclusions:
- Specific point mutations in the HBV core gene are common in chronically infected patients.
- PCR-RFLP offers an efficient method for detecting HBV viral mutants.
- This approach can aid in the rapid screening of large patient cohorts for HBV variants.
Abstract:
HBV chronic infection is an important health problem. The HBV core antigen carries several epitopes for T and B cell recognition and the immune response is crucial for determining the outcome of viral infection. Using PCR-RFLP several point mutations were detected in the HBV core ORF of HBV extracted from the serum of 140 chronically infected patients and 86 samples from another 37 patients followed-up in a longitudinal study. Mutations at position 2248 and 2147 (A3) and at 2038 (M2) were found most frequently. The wild type core genotype was found in about 50% of the samples. PCR-RFLP results were confirmed by direct sequencing of amplified products from HBV DNA present in chronically infected patients. The method is rapid and reliable and may be particularly useful for a rapid detection of viral mutants in a large number of patients.
