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

Area of Science:

• Hepatology
• Molecular Diagnostics
• Infectious Diseases

Background:

• Hepatitis B virus (HBV) infection remains a significant global health concern, with high mortality rates despite vaccination.
• Current antiviral treatments and diagnostic methods for chronic HBV infection have limitations in detecting early viral replication and managing complications.
• Existing serological tests and polymerase chain reaction (PCR) methods for HBV detection have drawbacks in speed, sensitivity, or accessibility.

Purpose of the Study:

• To develop and validate a rapid, sensitive, and portable loop-mediated isothermal amplification (LAMP) assay for detecting HBV DNA in plasma.
• To evaluate the performance characteristics of the developed LAMP assay compared to existing diagnostic methods.
• To assess the potential utility of the LAMP assay as a screening tool in resource-limited settings.

Main Methods:

• Development of a system-integrative, colorimetric loop-mediated isothermal amplification (LAMP) assay for HBV DNA detection.
• Optimization of amplification time and temperature (45 min at 62°C).
• Evaluation of assay specificity, sensitivity, and detection limit using 77 HBV-positive plasma samples.

Main Results:

• The optimized LAMP assay achieved a detection limit of 10 copies/µL.
• The assay demonstrated 100% specificity and 92.20% sensitivity.
• The entire detection process, including amplification, was completed in under 45 minutes.

Conclusions:

• The developed colorimetric LAMP assay is a sensitive, efficient, and reliable method for the rapid detection of HBV DNA.
• This portable assay shows significant potential for use as a screening test in areas with limited laboratory infrastructure and resources.
• The LAMP assay offers a promising alternative for early HBV detection, potentially improving patient management and outcomes.