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Serum HBV DNA detected by PCR in dot blot negative HBV chronic carriers with active liver disease
J Monjardino1, J Velosa, H C Thomas
1Academic Department of Medicine, St. Mary's Hospital Medical School, London, United Kingdom.
Insights
Polymerase chain reaction (PCR) detects low-level hepatitis B virus (HBV) DNA in chronic carriers with active liver disease, previously undetected by older methods. This finding supports HBV
Area of Science:
- Hepatology
- Virology
- Immunology
Background:
- Chronic hepatitis B virus (HBV) carriers with active liver disease often show undetectable serum HBV DNA using conventional methods like dot-blot hybridization.
- Understanding low-level HBV replication is crucial for elucidating disease pathogenesis and developing effective treatment strategies.
Purpose of the Study:
- To re-investigate HBV DNA levels in chronic carriers with active liver disease and previously undetected HBV DNA.
- To assess the sensitivity of polymerase chain reaction (PCR) for detecting low-level HBV DNA.
Main Methods:
- Serum samples from 49 chronic HBV carriers with active liver disease and undetected HBV DNA (dot-blot) were analyzed.
- Polymerase chain reaction (PCR) followed by Southern blot analysis was employed for sensitive HBV DNA detection.
- Patients were categorized into persistently HBeAg-negative and anti-HBe-positive groups.
Main Results:
- PCR detected HBV DNA in 87.5% (14/16) of HBeAg-negative and 81.8% (27/33) of anti-HBe-positive patients.
- HBV DNA detection sensitivity using PCR was significantly enhanced, reaching 10-50 virions/ml.
- Four of nine cases with undetected HBV DNA were positive for hepatitis delta virus (HDV) markers.
Conclusions:
- Low-level HBV replication is common in chronic HBV carriers with active liver disease, even when previously undetectable.
- PCR is a highly sensitive method for detecting low-level HBV DNA, essential for understanding HBV pathogenesis.
- Findings support the role of cytotoxic T lymphocyte-mediated immunopathogenesis and suggest combined antiviral and immunotherapeutic approaches.
Abstract:
A group of forty-nine HBV chronic carriers with histologically confirmed active liver disease and undetected serum HBV DNA by dot-blot hybridisation were re-investigated using the polymerase chain reaction (PCR) for amplification of serum DNA. The group comprised 16 persistently serum HBeAg-negative and thirty-three anti-HBe-positive patients. The use of PCR followed by Southern blot analysis has increased the sensitivity of HBV DNA detection to about 10-50 virions per ml of serum. Our results showed 14/16 (87.5%) of the HBeAg-positive group and 27/33 (81.8%) of the anti-HBe group to be positive for HBV DNA using PCR. Of the nine cases where HBV DNA was undetected four were positive for markers of hepatitis delta virus (HDV) infection. Demonstration of low level HBV replication associated with active liver disease in chronic HBV carriers where it was previously undetected meets a basic requirement for the proposed role of cytotoxic T lymphocyte-mediated immunopathogenesis in chronic hepatitis B and suggests a combined antiviral and immunotherapeutic approach to achieve eradication of the infection.