Optimization and Implementation of the Virus Extraction Method for Hepatitis E Virus Detection from Raw Pork Liver

Area of Science:

• Foodborne pathogens
• Virology
• Molecular detection

Background:

• Hepatitis E virus (HEV) is found in pork liver, posing risks to specific populations.
• Standardized methods for HEV detection in pork liver are lacking.
• Undercooked pork liver consumption is common in Asia.

Purpose of the Study:

• To optimize a virus extraction method for HEV from raw pork liver.
• To assess HEV prevalence in retail pork liver samples.
• To evaluate the public health implications of molecular detection results.

Main Methods:

• Compared various sample preparation protocols for HEV extraction.
• Utilized FastPrep™ homogenization with specific mechanical force parameters.
• Employed reverse transcription-quantitative polymerase chain reaction (RT-qPCR) for HEV detection.
• Performed RNase treatment to differentiate between infectious virus and RNA traces.

Main Results:

• An optimized homogenization protocol yielded satisfactory virus recovery rates (>1%).
• HEV genome was detected in 52% of 60 retail pork liver samples.
• RNase treatment reduced HEV prevalence to 33.3%, with positive samples showing high Ct values (>35).

Conclusions:

• The optimized method effectively extracts HEV from raw pork liver.
• High prevalence of HEV genome in pork liver warrants careful interpretation.
• Positive RT-qPCR signals may largely represent inactive viruses or RNA, necessitating caution in public health risk assessment.