Optimization of the Elution Buffer and Concentration Methods to Detect Hepatitis E Virus in Meat Using Nested Reverse Transcription-polymerase Chain Reaction and Real-time Reverse Transcription-polymerase Chain Reaction

Introduction: Hepatitis E virus (HEV) causing acute hepatitis through contaminated water or food is a major concern of public health in developing countries. The consumption of meat contaminated with HEV is an important transmission route.

Purpose: The purpose of this study was to optimize the elution and concentration methods for HEV in pork liver and to perform the comparative detection of nested RT-PCR and real-time RT-PCR for HEV in pork meat.

Methods: One gram and 10 g of swine liver was eluted using phosphate-buffered saline, threonine buffer, and glycine buffer. Polyethylene glycol (PEG) precipitation and ultrafiltration (UF) were used to concentrate HEV. Nested RT-PCR and real-time RT-PCR were compared for the detection of HEV in liver samples.

Results: When PBS elutes of 10 g liver samples were concentrated with UF, 6 (23.1%) out of 26 were HEV positive by real-time RT-PCR but all were negative for HEV by nested RT-PCR. PBS was the optimal buffer to elute HEV from 10 g liver sample. The combination of UF and real-time RT-PCR is the most sensitive detection method for HEV.

Significance: Compared with previous studies, this study developed the optimal protocol for the elution, concentration, and detection method for HEV in pork liver.