Development of Molecular-based Screening Test for Hepatitis B Virus in Human Plasma Samples

Abstract

Despite the HBV vaccine being routinely administered in many countries, the death rate remains significant. Antiviral medications are available for the treatment of HBV infection, but patients encounter various serious complications in cases of chronic HBV infection. The failure of serological tests to detect early viral replication prevents early treatment response. Recently, many studies have demonstrated significant advantages of the loop-mediated isothermal amplification (LAMP) assay over serological testing and PCR for the rapid detection of microbial pathogens. This study developed a rapid, sensitive, and portable system-integrative LAMP assay to detect Hepatitis B DNA in plasma samples. The final optimized assay was achieved with an amplification time of less than 45 minutes at 62°C. As a result of testing 77 HBV-positive plasma samples with known Cq values, the LAMP assay showed 100% specificity, 92, 20% sensitivity, and a detection limit of 10 copies/µL. Our results showed that the colorimetric LAMP assay is a sensitive, efficient, and highly reliable assay for the rapid detection of HBV and has the potential to be used as a screening test in areas where poor laboratory facilities and limited resources are available.