Multicentre collaborative trial evaluation of a method for detection of Hepatitis E Virus in pork liver, modified to contain an internal amplification control

Abstract

Demonstration that a detection method and its critical features is repeatable and reproducible is essential for its adoption as an international standard. An existing RT-qPCR-based method for detection of hepatitis E virus (HEV) in pig liver was modified to incorporate an internal amplification control, enabling a more accurate analysis with less control sample numbers. The method was subjected to interlaboratory trial involving seven laboratories from six European countries. Each laboratory tested eight samples at four artificial contamination levels: 0 genome copies (gc) HEV, 6 × 104 genome copies, 6 × 103 gc, and 6 × 102 gc per 25 mg liver. Trial sensitivity, or correct identification of positive samples, was 83.3%; the accordance was 82.2%, and the concordance was 69.6%. The positive predictive value was 93.8%. The trial specificity, or correct identification of uncontaminated samples, was 83.3%; the accordance was 66.7%, and the concordance was 70.0%. The negative predictive value was 62.5%. The internal amplification control (IAC) was detected in all samples except one. The results of the study are anticipated to assist in current standardization activities. The methodological advancement should aid in more accurate and reliable analysis, contributing to the broader goal of ensuring safer food supply chains.</p