Purpose: The objective of this study was to develop and validate a global ceeramTools®/bioMérieux detection method from samples to results in accordance with ISO/TS15216.
Methods: The performance criteria (evaluation of inhibition impact, specificity, limit of detection) of the complete solution was compared with the ones of the ISO/TS 15216-1 and 2. Protocols described specifically for each type of matrices (shellfish, soft fruits, leafy greens, bottled water and surface) included in the ISO/TS15216 were used for virus elution and concentration. Mengo virus VMC0 (ceeramTools®) was used as a viral process control. RNA extractions were performed using the bioMérieux NucliSens® MiniMag® DNA/RNA extraction platform. ceeramTools® molecular detection kits were used for identification and quantification of Mengo virus, norovirus GI and GII and hepatitis A virus. A ring trial study for each type was conducted with different laboratories.
Results: Whatever the category of samples, evaluation of inhibition leads to exactly the same results using either an external positive control EPC as described in ISO/TS 15216 or internal positive control IPC included in ceeramTools® detection kits for NoV GI, NoV GII and HAV targets. The same limit of detection was achieved (500 genome copies with IC > 95%) for all type of food samples (shellfish, soft fruits, leafy greens, bottled water and surface) with both methods. All the participants in the inter-laboratory study were able to reach the same limit of detection whatever the matrices.
Significance: For all sample categories included in the reference method, the complete solution and the reference method ISO/TS 15216 were concluded to be equivalent.