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Updated: Apr 6, 2026
Detection of Low Copy Number Integrated Viral DNA Formed by In Vitro Hepatitis B Infection
Published on: November 7, 2018
Usefulness of in-house PCR methods for hepatitis B virus DNA detection
Moyra Machado Portilho1, Marcia Leite Baptista2, Messias da Silva3
1Laboratory of Viral Hepatitis, Oswaldo Cruz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rio de Janeiro, Brazil.
Insights
In-house semi-nested PCR demonstrates good performance for Hepatitis B virus (HBV) DNA detection, showing adequate concordance with commercial assays. This makes it a viable alternative for HBV molecular diagnosis in resource-limited settings.
Area of Science:
- Molecular Biology
- Virology
- Clinical Diagnostics
Background:
- Hepatitis B virus (HBV) infection is a significant global health concern.
- Accurate and accessible HBV DNA detection is crucial for diagnosis and management.
- Commercial quantitative methods for HBV DNA detection can be expensive and inaccessible in low-resource settings.
Purpose of the Study:
- To evaluate the performance of three in-house PCR techniques for HBV DNA detection.
- To compare in-house methods with commercial quantitative assays.
- To assess the utility of in-house PCR for HBV diagnosis in resource-limited environments.
Main Methods:
- Three panels of HBsAg-reactive sera were tested using in-house qualitative PCR (core gene, pre-S/S gene, semi-nested PCR) and commercial assays (Cobas Amplicor HBV Monitor, Cobas TaqMan HBV).
- Performance was evaluated based on HBV DNA detection rates and concordance with commercial methods.
- Serial samples from HBV-infected individuals were analyzed to assess detection over time.
Main Results:
- In-house semi-nested PCR showed 90% concordance with Cobas Amplicor in panel I and 67.8% concordance with Cobas TaqMan in panel II.
- Other in-house PCR methods (core gene, pre-S/S gene) exhibited lower concordance rates.
- Semi-nested PCR detected HBV DNA in serial samples, though commercial assays sometimes provided detection for longer periods.
Conclusions:
- In-house semi-nested PCR offers adequate concordance with commercial methods for HBV DNA detection.
- It presents a potentially valuable and cost-effective alternative for molecular HBV diagnosis in settings with limited resources.
- Further validation may be needed for widespread adoption in low-resource laboratories.
Abstract:
The aim of the present study was to evaluate the performance of three in-house PCR techniques for HBV DNA detection and compare it with commercial quantitative methods to evaluate the usefulness of in-house methods for HBV diagnosis. Three panels of HBsAg reactive sera samples were evaluated: (i) 50 samples were examined using three methods for in-house qualitative PCR and the Cobas Amplicor HBV Monitor Assay; (ii) 87 samples were assayed using in-house semi-nested PCR and the Cobas TaqMan HBV test; (iii) 11 serial samples obtained from 2 HBV-infected individuals were assayed using the Cobas Amplicor HBV test and semi-nested PCR. In panel I, HBV DNA was detected in 44 samples using the Cobas Amplicor HBV test, 42 samples using semi-nested PCR (90% concordance with Cobas Amplicor), 22 samples using PCR for the core gene (63.6% concordance) and 29 samples using single-round PCR for the pre-S/S gene (75% concordance). In panel II, HBV DNA was quantified in 78 of the 87 HBsAg reactive samples using Cobas TaqMan but 52 samples using semi-nested PCR (67.8% concordance). HBV DNA was detected in serial samples until the 17th and 26th week after first donation using in-house semi-nested PCR and the Cobas Amplicor HBV test, respectively. In-house semi-nested PCR presented adequate concordance with commercial methods as an alternative method for HBV molecular diagnosis in low-resource settings.
