Molecular Detection of Hepatitis B Virus Among HBsAg Non-Reactive Blood of Donors

Main Article Content

Habibat Bello
AbdulKadir Bashir
Babalola Oladayo Musa
Ibrahim Damilare Boyenle
Ashiru Mojeed Ayoola
R. A. Sulyman


Despite all blood donations being tested routinely for HBsAg as a clinical marker of transmissible HBV, cases of post-transfusion hepatitis B virus infection are still being reported because molecular studies using Polymerase Chain Reaction (PCR) are not routinely available for Transfusion Transmissible Infection (TTIs) testing. In this study, we sought to use the PCR technique to re-screen donated blood that had already been proven to be HBsAg non-reactive with rapid diagnostic testing and ELISA. One hundred and eighty-five samples were obtained from a proportion of the blood deposited at the blood bank of the Federal Medical Center, Birnin-Kebbi, Nigeria. Socio-demographic parameters such as age group, status, ethnicity, occupation, and group PCV were obtained from donors' records. Nested PCR was employed to detect HBV DNA. Furthermore, genotyping was performed to determine HBV genotypes in the positive samples using PCR with genotype-specific primers. Of the 185 donors, it was observed that five (2.7%) of the population were positive for HBV. HBV is more common among people aged 18–30, singles, Hausa, and self-employed. In addition, the five positive samples were of genotype E. This study suggests the need to complement antibody-based tests with DNA testing for effective HBV screening and consequent safe transfusion.

Keywords: Hepatitis B Virus; HBsAg; PCR; DNA testing


Download data is not yet available.


Metrics Loading ...

Article Details

How to Cite
Bello, H., Bashir, A., Musa, B. O., Boyenle, I. D., Ayoola, A. M., & Sulyman, R. A. (2022). Molecular Detection of Hepatitis B Virus Among HBsAg Non-Reactive Blood of Donors. Fountain Journal of Natural and Applied Sciences, 11(2).


Adekeye, A. M., Chukwuedo, A. A., Zhakom, P. N., & Yakubu, R. S. (2013). Prevalence of hepatitis B and C among blood donors in Jos South LGA, Plateau State, Nigeria. Asian Journal of Medical Sciences, 5(5), 101–104.

Adoga, M. P., Gyar, S. D., Pechulano, S., Bashayi, O. D., Emiasegen, S. E., Zungwe, T., Iperepolu, O. H., Agupugo, C., & Agwale, S. M. (2010). Hepatitis B virus infections in apparently healthy urban Nigerians: data from pre-vaccination tests. The Journal of Infection in Developing Countries, 4(06), 397–400.

Ahmad, G., Khan, M. A., Abbas, S., Athar, A., Khan, B. S., & Aslam, M. S. (2019). Automated diagnosis of hepatitis b using multilayer mamdani fuzzy inference system. Journal of Healthcare Engineering, 2019.

Amadei, B., Urbani, S., Cazaly, A., Fisicaro, P., Zerbini, A., Ahmed, P., Missale, G., Ferrari, C., & Khakoo, S. I. (2010). Activation of natural killer cells during acute infection with hepatitis C virus. Gastroenterology, 138(4), 1536–1545.

Antar, W., El?Shokry, M. H., Abd El Hamid, W. A., & Helmy, M. F. (2010). Significance of detecting anti?HBc among Egyptian male blood donors negative for HBsAg. Transfusion Medicine, 20(6), 409–413.

Cohen Stuart, J. W. T., Velema, M., Schuurman, R., Boucher, C. A. B., & Hoepelman, A. I. M. (2009). Occult hepatitis B in persons infected with HIV is associated with low CD4 counts and resolves during antiretroviral therapy. Journal of Medical Virology, 81(3), 441–445.

Di Lello, F. A., Macias, J., Cifuentes, C. C., Vargas, J., Palomares, J. C., & Pineda, J. A. (2012). Low prevalence of occult HBV infection among HIV-infected patients in Southern Spain. Enfermedades Infecciosas y Microbiologia Clinica, 30(6), 312–314.

Dongdem, A. Z., Dzodzomenyo, M., Asmah, R. H., Nyarko, K. M., Nortey, P., Agyei, A., Adjei, D. N., Kenu, E., & Adjei, A. A. (2016). Hepatitis B virus genotypes among chronic hepatitis B patients reporting at Korle-Bu teaching hospital, Accra, Ghana. The Pan African Medical Journal, 25(Suppl 1).

Eke, A. C., Eke, U. A., Okafor, C. I., Ezebialu, I. U., & Ogbuagu, C. (2011). Prevalence, correlates and pattern of hepatitis B surface antigen in a low resource setting. Virology Journal, 8(1), 1–8.

El?Zayadi, A., Ibrahim, E. H., Badran, H. M., Saeid, A., Moneib, N. A., Shemis, M. A., Abdel?Sattar, R. M., Ahmady, A. M., & El?Nakeeb, A. (2008). Anti?HBc screening in Egyptian blood donors reduces the risk of hepatitis B virus transmission. Transfusion Medicine, 18(1), 55–61.

Elmaghloub, R., Elbahrawy, A., Didamony, G. El, Elwassief, A., Saied Mohammad, A.-G., Alashker, A., Zedan, H., Abdallah, A. M., Hemidah, M. H., Elmestikawy, A., Fayoumei, M. El, Shahba, H., Gawish, A., Morsy, M. H., Hashim, A., Abdelbaseer, M. A., Ueda, Y., Chiba, T., & Abdelhafeez, H. (2017). Hepatitis B Virus Genotype E Infection among Egyptian Health Care Workers. Journal of Translational Internal Medicine, 5(2), 100–105.

Faleye, T. O. C., Adewumi, M. O., Ifeorah, I. M., Omoruyi, E. C., Bakarey, S. A., Akere, A., Awokunle, F., Ajibola, H. O., Makanjuola, D. O., & Adeniji, J. A. (2015). Detection of hepatitis B virus isolates with mutations associated with immune escape mutants among pregnant women in Ibadan, southwestern Nigeria. Springerplus, 4(1), 1–8.

Fang, Z., Harrison, T. J., Yang, J., Chen, Q., Wang, X., & Mo, J. (2012). Prevalence of hepatitis B virus infection in a highly endemic area of southern China after catch?up immunization. Journal of Medical Virology, 84(6), 878–884.

Ifeorah, I. M., Bakarey, A. S., Adewumi, M. O., Faleye, T. O. C., Akere, A., Omoruyi, C. E., Ogunwale, A. O., Uttah, C. C., Oketade, M. A., & Adeniji, J. A. (2017). Patterns of serologic markers of hepatitis B virus infection and the risk of transmission among pregnant women in southwestern Nigeria. Journal of Immunoassay and Immunochemistry, 38(6), 639–651.

Jafarzadeh, A., Arababadi, M. K., & Pourazar, M. M. A. (2008). Occult hepatitis B virus infection among blood donors with antibodies to hepatitis B core antigen. Acta Medica Iranica, 27–32.

Janahi, E. M., Ilyas, Z., Al-Othman, S., Darwish, A., Sanad, S. J., Al-musaifer, B., Al-Mannai, M., Golbahar, J., & Perna, S. (2019). Hepatitis B virus genotypes in the kingdom of Bahrain: Prevalence, gender distribution and impact on hepatic biomarkers. Medicina (Lithuania), 55(10).

Kurbanov, F., Tanaka, Y., & Mizokami, M. (2010). Geographical and genetic diversity of the human hepatitis B virus. Hepatology Research: The Official Journal of the Japan Society of Hepatology, 40(1), 14–30.

Liang, S.-H., Chen, T.-J., Lee, S. S.-J., Tseng, F.-C., Huang, C.-K., Lai, C.-H., Chiou, C.-P., Wang, J.-L., Chung, H.-C., Lin, J.-N., & others. (2010). Risk factors of isolated antibody against core antigen of hepatitis B virus: association with HIV infection and age but not hepatitis C virus infection. JAIDS Journal of Acquired Immune Deficiency Syndromes, 54(2), 122–128.

Matlou, M. K., Gaelejwe, L. R., Musyoki, A. M., Rakgole, J. N., Selabe, S. G., & Amponsah-Dacosta, E. (2019). A novel hepatitis B virus recombinant genotype D4/E identified in a South African population. Heliyon, 5(4), e01477.

Neau, D., Winnock, M., Jouvencel, A.-C., Faure, M., Castéra, L., Legrand, E., Lacoste, D., Ragnaud, J.-M., Dupon, M., Fleury, H., & others. (2005). Occult hepatitis B virus infection in HIV-infected patients with isolated antibodies to hepatitis B core antigen: Aquitaine cohort, 2002--2003. Clinical Infectious Diseases, 40(5), 750–753.

Nna, E., Mbamalu, C., & Ekejindu, I. (2014). Occult hepatitis B viral infection among blood donors in South–Eastern Nigeria. Pathogens and Global Health, 108(5), 223–228.

O’Brien, S. F., Fearon, M. A., Yi, Q., Fan, W., Scalia, V., Muntz, I. R., & Vamvakas, E. C. (2007). Hepatitis B virus DNA–positive, hepatitis B surface antigen–negative blood donations intercepted by anti?hepatitis B core antigen testing: the Canadian Blood Services experience. Transfusion, 47(10), 1809–1815.

Odemuyiwa, S. O., Mulders, M. N., Oyedele, O. I., Ola, S. O., Odaibo, G. N., Olaleye, D. O., & Muller, C. P. (2001). Phylogenetic analysis of new hepatitis B virus isolates from Nigeria supports endemicity of genotype E in West Africa. Journal of Medical Virology, 65(3), 463–469.

Okonko, S. F. A., Alli, J. A., Ojezele, M. O., Udeze, A. O., Nwanze, J. C., & Adewale, O. (2010). Seroprevalence of HBsAg Antigenemia among Patients in Abeokuta. South Western Nigeria, 10(2), 40–46.

Oladele, O. O., Adesina, T. B., Omolola, Z. T., Sunday, T. S., Shabir, B., & Raza, A. (2013). Prevalence of HBsAg and HIV among blood donors in Osogbo, Osun State, Nigeria. International Research Journal of Medicine and Medical Sciences, 1(3), 68–71.

Olotu, A. A., Oyelese, A. O., Salawu, L., Audu, R. A., Okwuraiwe, A. P., & Aboderin, A. O. (2016). Occult Hepatitis B virus infection in previously screened, blood donors in Ile-Ife, Nigeria: implications for blood transfusion and stem cell transplantation. Virology Journal, 13(1), 1–8.

Oluyinka, O. O., Tong, H. Van, Bui Tien, S., Fagbami, A. H., Adekanle, O., Ojurongbe, O., Bock, C.-T., Kremsner, P. G., & Velavan, T. P. (2015). Occult hepatitis B virus infection in Nigerian blood donors and hepatitis B virus transmission risks. PloS One, 10(7), e0131912.

Opaleye, O. O., Fagbami, A. H., & Bolaji, O. S. (2014). Occult hepatitis B virus infection in fully screened blood units in Nigeria. International Journal of Infectious Diseases, 21, 455.

Osuji, A., Agbakoba, N. R., Ifeanyichukwu, M. O., & Tatfeng, M. (2018). Occult hepatitis B virus infection among blood donors at two teaching hospitals in Nigeria: implications for blood transfusion. Amer. J. Infect. Dis. Microbiol, 6(1), 16–25.

OSUJI, A. I., Duru, G. C., & Kinjir, H. (2021). Detection of Hepatitis C virus antibodies among healthy blood donors at a tertiary hospital in Nigeria. Microbes and Infectious Diseases, 2(2), 252–259.

Rajalakshmi, S. (2017). Different Types of PCR Techniques and Its Applications. International Journal of Pharmaceutical, Chemical & Biological Sciences, 7(3).

Salawu, L., Bolarinwa, R. A., Adegunloye, A. B., & Muraina, H. A. (2010). HBsAg, anti-HCV, anti-HIV and VDRL in blood donors: Prevalence and trends in the last three and a half years in a tertiary health care facility in Ile-Ife, Nigeria. International Journal of Medicine and Medical Sciences, 2(11), 335–341.

Salih, E. Y., Saeed, W. S. E., Satti, M. M. H., Abbas, G., Suleiman, I. A., & Khalil, E. A. G. (2018). Hepatitis B virus genotypic patterns in Sudan reflect population movements. Glob. J. Virol. Immunol, 3, 176–180.

Singh, J., Birbian, N., Sinha, S., & Goswami, A. (2014). A critical review on PCR, its types and applications. Int J Adv Res Biol Sci, 1(7), 65–80.

Tramuto, F., Maida, C. M., Colomba, G. M. E., Di Carlo, P., & Vitale, F. (2013). Prevalence of occult hepatitis B virus infection in a cohort of HIV-positive patients resident in Sicily, Italy. BioMed Research International, 2013, 859583.

WHO. (2000). Hepatitis B. Fact sheet no. 204. World Health Organization Geneva.

Yakubu, A., Sahabi, D. M., Saidu, Y., & Magaji, U. F. (2016). Prevalence of HBsAg among prospective blood donors and pregnant women in Kebbi State, Nigeria. Nigerian Journal of Basic and Applied Sciences, 24(1), 57–63.