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Updated: Jul 5, 2026
Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
Diagnosing different stages of hepatitis B infection using a competitive polymerase chain reaction assay
H Changotra1, A Dwivedi, A K Nayyar
1Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130, USA.
Insights
Serum HBV DNA levels help stage hepatitis B virus (HBV) infection. A 2.5 x 10^4 copies/mL cutoff optimally distinguishes inactive carriers from HBeAg-negative chronic HBV patients in India.
Area of Science:
- Hepatology
- Virology
- Molecular Diagnostics
Background:
- Hepatitis B virus (HBV) infection stages correlate with serum HBV DNA levels.
- Distinguishing inactive carriers from chronic HBV patients is crucial for management.
Purpose of the Study:
- To investigate serum HBV DNA levels in inactive carriers and chronic HBV (CHB) patients.
- To define a cutoff value for differentiating inactive carriers from HBeAg-negative CHB patients in the Indian population.
Main Methods:
- Analysis of 140 samples: 42 inactive HBsAg carriers and 98 CHB patients (53 HBeAg-positive, 45 HBeAg-negative).
- Serum HBV DNA quantification using an in-house competitive polymerase chain reaction (cPCR) assay.
Main Results:
- HBeAg-positive patients had significantly higher HBV DNA loads than HBeAg-negative patients (median 1.25 x 10^8 vs. 2.30 x 10^5 copies/mL).
- HBeAg-negative CHB patients showed significantly higher HBV DNA levels than inactive carriers (median 2.30 x 10^5 vs. 4.28 x 10^3 copies/mL).
- A cutoff of 2.5 x 10^4 copies/mL demonstrated 75.6% sensitivity and 78.6% specificity for discriminating HBeAg-negative CHB from inactive carriers.
Conclusions:
- A serum HBV DNA cutoff of 2.5 x 10^4 copies/mL is proposed for classifying inactive carriers and HBeAg-negative CHB patients.
- This cutoff aids in better management of HBV infection despite overlapping DNA levels.
- Accurate staging is essential for effective therapeutic strategies in hepatitis B.
Purpose:
Different stages of hepatitis B virus (HBV) infection can be defined by serum HBV DNA levels. This study attempts to (1) investigate serum HBV DNA levels in inactive carriers and patients with chronic HBV (CHB) infection and (2) define cut-off value between inactive carriers and HBeAg (precore antigen of HBV) negative CHB patients in Indian population.
Methods:
One hundred and forty samples encompassing 42 inactive HBsAg carriers and 98 CHB patients (53 HBeAg-positive and 45 HBeAg-negative) were analysed. Serum HBV DNA levels were determined employing an in-house competitive polymerase chain reaction (cPCR) assay.
Results:
The HBeAg-positive patients were found to have the maximum median HBV DNA load, which was significantly higher than the HBeAg-negative ones (median; 1.25 x 10(8) vs. 2.30 x 10(5) copies/mL ; P<0.05). Interestingly, the latter group has significantly higher HBV DNA levels than the inactive carriers (median; 2.30 x 10(5) vs. 4.28 x 10(3) copies/mL; P<0.05). The 2.5 x 10(4) copies/ml HBV DNA levels were optimal for discriminating CHB patients (HBeAg-negative) from inactive carriers with 75.6 and 78.6% sensitivity and specificity, respectively.
Conclusions:
Despite the extensive overlapping of HBV DNA levels in inactive carriers and HBeAg negative CHB patients, 2.5 x 10(4) copies/mL is the most favourable cut-off value to classify these individuals and would be imperative in the better management of this dreadful disease.
