Simultaneous detection of HBV and HCV by multiplex PCR normalization

Area of Science:

• Molecular Biology
• Virology
• Diagnostic Assays

Background:

• Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) are significant global health concerns.
• Simultaneous detection methods are crucial for efficient diagnosis and management.
• Existing multiplex PCR techniques may face challenges with multiplex factor interference.

Purpose of the Study:

• To develop and validate a normalized multiplex PCR method for simultaneous HBV and HCV detection.
• To optimize the method by comparing nucleic acid extraction techniques and key PCR factors.

Main Methods:

• A two-round PCR amplification using primers with a joint sequence, followed by a final amplification using the joint sequence.
• Normalization to a single PCR system to mitigate multiplex factor interference.
• Orthogonal design for optimizing six key factors and evaluation of four nucleic acid extraction methods.

Main Results:

• The developed method demonstrated good sensitivity and specificity for detecting HBV and HCV.
• Diagnostic indices and efficiency were evaluated across different patient sample types.
• Validation was performed on twenty serum samples, confirming the method's validity and authenticity.

Conclusions:

• The normalized multiplex PCR method is effective for simultaneous amplification of multiple viral targets.
• The technique is practical, accurate, and suitable for the prevention and control of HBV and HCV infections.
• This approach holds promise for broader applications in detecting multiple genes from various sources.