Current Trends in the Laboratory Diagnosis of Hepatitis B Infections
Article Sidebar
Main Article Content
Abstract
Viral hepatitis remains a major global public health challenge, accounting for approximately 1.4 million deaths annually.Hepatitis B virus (HBV) infection alone contributes substantially to this burden, particularly in low- and middle-income countries where access to timely diagnosis and treatment remains limited. Despite the availability of effective vaccines and antiviral therapies, an estimated 90% of individuals living with HBV worldwide remain unaware of their infection status, perpetuating transmission and increasing the risk of long-term liver complications. This review examines current trends in the laboratory diagnosis of HBV infection, with emphasis on both conventional and emerging diagnostic technologies. A systematic literature review was conducted using PubMed, Scopus, and Google Scholar to identify relevant studies published between 2010 and 2025 on HBV diagnostic methodologies. Traditional diagnostic approaches, including serological markers, biochemical
liver function tests, and imaging techniques, remain foundational for assessing infection status, immune response, and liver disease progression. However, molecular diagnostics have become central to HBV management, with quantitative detection of HBV DNA and RNA now regarded as the gold standard for viral load monitoring and therapeutic decision-making. Advances in molecular diagnostics span multiple generations, including real-time quantitative polymerase chain reaction (qPCR), isothermal amplification technologies, HBV RNA quantification, and next-generation sequencing (NGS). These innovations offer enhanced sensitivity and specificity, improved disease monitoring, rapid turnaround times, high-throughput capabilities, and expanded applications such as genotyping and antiviral resistance detection. Despite these advancements, significant challenges persist, notably high costs, infrastructure demands, lack of standardization, and limited accessibility in underserved populations. Achieving the World Health Organization’s 2030 hepatitis B elimination targets will require expanding access to affordable, accurate, and simplified diagnostic tools alongside strengthened health systems, workforce training, and public
awareness initiatives. Continued technological innovation and global collaboration remain essential to reducing diagnostic gaps and improving outcomes for millions affected by HBV worldwide.
Downloads
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
Authors hold the copyright of all published articles except otherwise stated.
References
Alekseenko, A., Ignatyev, S., Saenko, S., & Zaitsev, D. (2022). Clinical implementation of next-generation sequencing: Challenges and opportunities. Clinical Chemistry and Laboratory Medicine, 60(4), 567–578. https://doi.org/10.1515/cclm-2021-1023
Alsudaney, A. A., Alshammari, F. D., & Alenzi, A. M. (2025). Advances in PCR-based mutation detection techniques in viral diagnostics. Journal of Molecular Diagnostics, 27(1), https://doi.org/10.1016/j.jmoldx.2024.09.003 12–25.
Bong, J. H., Kim, S. U., Park, J. Y., & Ahn, S. H. (2024). Clinical applications of serum hepatitis B virus RNA quantification. Journal of Hepatology, 80(2), 402 414. https://doi.org/10.1016/j.jhep.2023.10.012.
Boonstra, A., & Sari, Y. (2025). Serum HBV RNA as a surrogate marker for cccDNA activity. Hepatology International, 19(1), 1–11. https://doi.org/10.1007/s12072-024-10567-9
Börsch-Supan, A., Jansen, L., & Urban, S. (2021). Hepatitis B virus RNA: Emerging biomarker for treatment monitoring. Gut, Table 2: Comparison of Molecular Techniques for HBV Detection 70(2), 384–392. https://doi.org/10.1136/gutjnl-2020-321468
Buti, M., Marcos-Fosch, C., & Esteban, R. (2021). Occult hepatitis B infection: Clinical implications. Liver International, 41(4), 697–708. https://doi.org/10.1111/liv.14759
Centers for Disease Control and Prevention. (2023). Hepatitis B questions and answers for health professionals. https://www.cdc.gov/hepatitis/hbv
Chen, Z., Li, Y., & Liu, J. (2019). Next-generation sequencing in viral diagnostics. Virology Journal, 16, Article 124. https://doi.org/10.1186/s12985-019-1228-6
Cholongitas, E., Papatheodoridis, G. V., & Vassiliadis, T. (2018). Clinical relevance of isolated anti-HBc positivity. Journal of Viral Hepatitis, 25(6), 614–620. https://doi.org/10.1111/jvh.12866
Chook, J. B., Ong, S. H., & Lee, C. K. (2015). Digital PCR and isothermal amplification technologies. Clinical Chemistry, 61(6), 867–876. https://doi.org/10.1373/clinchem.2014.236760 Compton, J. (2018). Isothermal nucleic acid amplification technologies. Nature Methods, 15(9), 671–673. https://doi.org/10.1038/s41592-018-0118-0
Cooreman, M. P., Van Der Auwera, G., & Vercauteren, J. (2021). Comparison of transcription-mediated amplification and real-time PCR for HBV viral load. Journal of Clinical Virology, 136, 104738. https://doi.org/10.1016/j.jcv.2021.104738
Craw, P., & Balachandran, W. (2012). Isothermal nucleic acid amplification technologies. Lab on a Chip, 12(14), 2469–2486. https://doi.org/10.1039/c2lc40100b De Clercq, E., & Li, G. (2010). Approved antiviral drugs over the past 50 years. Clinical Microbiology Reviews, 29(3), 695–747. https://doi.org/10.1128/CMR.00102 15
Dini, G., Toletone, A., & Durando, P. (2017). Hepatitis B vaccination and antibody persistence. Human Vaccines & Immunotherapeutics, 13(1), 124–130. https://doi.org/10.1080/21645515.2016.1239661
Bong, J. H., Kim, S. U., Park, J. Y., & Ahn, S. H. (2024). Clinical applications of serum hepatitis B virus RNA quantification. Journal of Hepatology, 80(2), 402 414. https://doi.org/10.1016/j.jhep.2023.10.012
Boonstra, A., & Sari, Y. (2025). Serum HBV RNA as a surrogate marker for cccDNA activity. Hepatology International, 19(1), 1–11. https://doi.org/10.1007/s12072-024-10567-9
Buti, M., Marcos-Fosch, C., & Esteban, R. (2021). Occult hepatitis B infection: Clinical implications. Liver International, 41(4), 697–708. https://doi.org/10.1111/liv.14759 Carulli, G. (2015). Real-time PCR applications in clinical virology. Journal of Clinical Laboratory Analysis, 29(3), 189–201. https://doi.org/10.1002/jcla.21765 Centers for Disease Control and Prevention. (2005). A comprehensive immunization strategy to eliminate transmission of hepatitis B virus infection. CDC.
Espy, M. J., Uhl, J. R., Sloan, L. M., & Smith, T. F. (2014). Real-time PCR in clinical microbiology. Clinical Microbiology Reviews, 19(1), 165–256. https://doi.org/10.1128/CMR.19.1.165-256.2006 European Association for the Study of the Liver. (2017). EASL 2017 clinical practice guidelines on hepatitis B virus infection. Journal of Hepatology, 67(2), 370 398. https://doi.org/10.1016/j.jhep.2017.03.021
Fakruddin, M., Mazumdar, R. M., Chowdhury, A., & Mannan, K. S. (2012). Nucleic acid amplification methods. Bangladesh Journal of Medical Science, 11(3), 195 205.
Ganczak, M., Dmytrzyk-Daniłów, G., & Korzeń, M. (2019). Isolated anti-HBc positivity. BMC Infectious Diseases, 19, Article 1017. https://doi.org/10.1186/s12879-019-4597-4
Grüner, N., Stambouli, O., & Ross, R. S. (2015). Dried blood spots for viral diagnostics. Journal of Clinical Virology, 72, 92–97. https://doi.org/10.1016/j.jcv.2015.09.012 Haddad-Boubaker,
S., Mbarek, H., & Yassine, H. M. (2023). Molecular diagnostics in hepatitis B management. Frontiers in Microbiology, 14, 1184729. https://doi.org/10.3389/fmicb.2023.1184729 Jefferies,
M., Rauff, B., Rashid, H., Lam, T., & Rafiq, S. (2018). Update on global epidemiology of viral hepatitis. World Journal of Gastroenterology, 24(9), 991–1003. https://doi.org/10.3748/wjg.v24.i9.991
Kacian, D. L., & Fultz, T. J. (2016). Transcription-mediated amplification. Clinical Chemistry, 62(6), 865–875. https://doi.org/10.1373/clinchem.2015.250563
Khan, A., Shah, M., & Iqbal, M. (2025). Digital PCR in viral diagnostics. Journal of Clinical Virology, 162, 105382. https://doi.org/10.1016/j.jcv.2024.105382
Liaw, Y. F., & Chu, C. M. (2009). Hepatitis B virus infection. The Lancet, 373(9663), 582–592. https://doi.org/10.1016/S0140-6736(09)60207-5
Liu, Y., Zhang, S., Chen, J., & Lu, F. (2020). Molecular virology of hepatitis B virus. World Journal of Gastroenterology, 26(6), 537–549. https://doi.org/10.3748/wjg.v26.i6.537
Lu, Y., Mo, Q., & Wang, J. (2006). Quantitative real-time PCR for HBV DNA. Journal of Virological Methods, 132(1–2), 41–46. https://doi.org/10.1016/j.jviromet.2005.08.020
Malagnino, V., Fofana, D. B., & Puro, V. (2018). Occult hepatitis B infection. Infectious Disease Reports, 10(1), 7484. https://doi.org/10.4081/idr.2018.7484
Milbury, C. A., Li, J., & Makrigiorgos, G. M. (2011). COLD PCR enrichment of rare mutations. Nature Methods, 8(2), 153–159. https://doi.org/10.1038/nmeth.1553 Nguyen, M. H., Wong, G., & Gane, E. (2020). Hepatitis B virus RNA: Clinical relevance. Hepatology, 72(1), 397–408. https://doi.org/10.1002/hep.31074
Notomi, T., Okayama, H., & Masubuchi, H. (2000). Loop mediated isothermal amplification of DNA. Nucleic Acids Research, 28(12), E63. https://doi.org/10.1093/nar/28.12.e63
Obiomah, P., Amilo, G. I., & Ndulue, J. (2020). HBV DNA quantification in Nigeria. African Journal of Laboratory Medicine, 9(1), 1023. https://doi.org/10.4102/ajlm.v9i1.1023 Odimayo,
M. S., Alabi, B., Adebimpe, W., Nwadioha, S., Olatunji, O., & Ogedengbe, B., (2022). Treatment Penetration and Correlates of Diagnostic Parameters among Hepatitis B Seropositive individuals in Ondo State, Nigeria. Western Journal of Medical and Biomedical Sciences. 2022; 3(1), 107-113. https//doi.org/10.5281/zenodo.5907250.
Okda, F., Kamal, M., & El-Sayed, M. (2016). Performance of rapid tests for HBsAg. Journal of Medical Virology, 88(2), 340–345. https://doi.org/10.1002/jmv.24312
Peeling, R. W., & Mabey, D. (2010). Point-of-care diagnostics for infectious diseases. Nature Reviews Microbiology, 8(12), 867–875. https://doi.org/10.1038/nrmicro2456 Roman, S. (2018). Occult hepatitis B virus infection. World Journal of Gastroenterology, 24(25), 2747–2759. https://doi.org/10.3748/wjg.v24.i25.2747
Seto, W. K., Lo, Y. R., & Pawlotsky, J. M. (2015). Chronic hepatitis B management. Journal of Hepatology, 63(1), S1–S36. https://doi.org/10.1016/j.jhep.2015.07.001
Spearman, C. W., Dusheiko, G. M., & Sonderup, M. W. (2023). Novel biomarkers in hepatitis B. Hepatology International, 17(3), 563–575. https://doi.org/10.1007/s12072-023-10459-3 Srivastava, S., & Prasad, R. (2023). Isothermal amplification technologies. Biosensors, 13(2), 196. https://doi.org/10.3390/bios13020196
Terrault, N. A., Lok, A. S., & McMahon, B. J. (2018). Update on prevention and treatment of hepatitis B. Hepatology, 67(4), https://doi.org/10.1002/hep.29800 1560–1599.
Tsuge, M., Murakami, E., & Imamura, M. (2019). Ultra sensitive HBsAg Gastroenterology, assays. 54(9), Journal of 737–747. https://doi.org/10.1007/s00535-019-01583-7
Vainionpää, R., & Leinikki, P. (2012). Diagnostic techniques in viral hepatitis. Scandinavian Journal of Clinical and Laboratory Investigation, 72(sup243), 31–36. https://doi.org/10.3109/00365513.2012.682893
Wang, J., Shen, T., & Huang, X. (2021). Serum HBV RNA as a novel biomarker. Clinical Gastroenterology and Hepatology, 19(4), 756–765. https://doi.org/10.1016/j.cgh.2020.04.052
World Health Organization. (2017). Guidelines on hepatitis B and C testing. WHO. World Health Organization. (2022). Global hepatitis report 2022. WHO.