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Updated: May 18, 2026
Real-Time Polymerase Chain Reaction-Based Detection and Quantification of Hepatitis B Virus DNA
Published on: December 15, 2023
The detection of occult HBV infection in patients with HBsAg negative pattern by real-time PCR method
Sena Izmirli1, Deniz Gozde Celik, Pelin Yuksel
1Microbiology and Clinical Microbiology Department, Cerrahpasa Faculty of Medicine, Istanbul University, Istanbul, Turkey.
Aim:
Diagnostic problems may be encountered in Hepatitis B virus (HBV) infections by serological tests and HBV DNA can be detectable in plasma and liver tissue while the HBsAg test is negative. This situation can be defined as occult or isolated Anti-HBc infections. Occult HBV infections may be divided into two categories by using hepatitis markers. One of them being that all hepatitis markers are negative and the other situation is having Anti-HBc +/- and Anti-HBs+patterns. These situations can be seen in isolated Anti-HBc cases.
Method:
In this study, we aimed to detect the ratio of occult HBV infections by investigating HBV DNA in four different groups. These groups are: (1) 20 isolated Anti-HBc positive individuals, (2) 23 individuals naturally immune to HBV infection, (3) 20 individuals with seronegative hepatitis markers and high ALT levels, and (4) 23 vaccinated individuals against HBV. In order to detect HBV DNA the real-time PCR kit (QIAGEN, Artus HBV RG PCR Kit, Germany) with high analytical sensitivity (≤3.8IU/ml) was used.
Results:
The reliability of the molecular methods was assessed by increasing the quantitation standards of internal, external and also positive controls. No HBV DNA was detected in any of the 86 individuals consisting of four study groups.
Conclusion:
In conclusion, we did not detect occult HBV infection in our four study groups by using a high sensitivity real-time (RT) PCR method, while occult HBV infections with various frequencies were detected in other large, serial international studies in which highly sensitive analytical molecular methods were used. Although we also used a high standard molecular kit to detect occult HBV infections, we suggest that the reason for the absence of detection of occult HBV infections may be due to the small number of cases included in this study. However, it was assumed that the use of a nucleic acid amplification technology (NAT) with high analytical sensitivity in blood banks to prevent HBV transmission by blood transfusion is controversial due to both costs and diagnostic efficacy and for this reason we suggest that it will be useful to perform large serial studies regarding occult HBV infections in the future.
