A molecular detection method for HBV pgRNA without RNA extraction based on nucleic acid hybridization

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Virology

Background:

• Early diagnosis of chronic hepatitis B (CHB) is crucial for patient outcomes.
• Current methods often require RNA extraction and purification, delaying diagnosis.
• Direct detection of viral RNA offers a potential simplification.

Purpose of the Study:

• To develop and validate a novel assay for direct detection of HBV pgRNA.
• To establish an efficient and high-throughput method for CHB screening.

Main Methods:

• A sandwich RNA hybridization assay was designed using oligonucleotide probes targeting conserved regions of HBV pgRNA.
• A hybridization cascade reaction was employed on a solid surface.
• The assay performance was evaluated using serum samples from CHB patients and healthy individuals.

Main Results:

• Optimal conditions were determined for probe concentrations, coating, and incubation times.
• The assay achieved a sensitivity of 91.47% and a specificity of 90.63%.
• The method directly detects HBV pgRNA, eliminating the need for RNA extraction.

Conclusions:

• The developed assay is simple, rapid, and high-throughput.
• This method is suitable for active CHB infection screening.
• Direct detection of HBV pgRNA facilitates early diagnosis and management.