Inter-laboratory validation study of two immunochemical methods for detection of processed ruminant proteins

In order to facilitate safe re-introduction of non-ruminant processed animal proteins (PAPs) in aqua feed, two immunoassays have been tested in an interlaboratory study for their capability to detect ruminant PAPs processed under European conditions. The sensitivity of the MELISA-TEK assay was improved by applying a specific extraction kit. Six approved blank pork and poultry samples were adulterated to produce 15 samples spiked at 0.5%, 1.0% and 2.0% with ruminant material, sterilised at either 133 °C or 137 °C. Fourteen participants investigated the 6 blanks and 15 spiked samples, making 21 samples for the final test. For both assays specificity and sensitivity were at 97% or higher. Concordance and accordance were higher than 95% with one exception. The results indicate that both assays provided correct results at 0.5% and higher for the detecting ruminant PAPs (sterilised at 133 °C) in non-ruminant PAPs. Given the 2% upper limit of ruminant PAPs in non-ruminant PAPs for avoiding an increase in BSE incidents, these methods are fit for monitoring non-ruminant PAPs intended for aqua feed.