The test detects DNA of crustaceans (Crustacea) according to directive (EC) 1169/2011 qualitatively and / or quantitatively.
For the quantitative determination the use of the laboratory reference material SureFood® QUANTARD Allergen 40 (Art. No. S3301) containing 40 mg crustaceans / kg food sample and a detection system including a standard series is required.
The real-time PCR assay can be performed with commonly used real-time PCR instruments, equipped for detection of two fluorescence emissions at the channels FAM and VIC/HEX at the same time. The technical validation of instruments was performed on Roche LightCycler® 480 II, Roche cobas® z 480 Analyzer, Qiagen Rotor-Gene Q, Applied Biosystems 7500, Bio-Rad CFX96, Bio Molecular Systems MIC, Agilent AriaDx and Agilent Mx3005P.
|Test format||100 reactions|
|Sample preparation||For DNA-preparation the use of SureFood® PREP Advanced, protocol 1 is recommended.|
|Detection limit||The method is validated for quantitative determination of 1 mg to 400 mg allergenic substance / kg food sample using the reference material SureFood® QUANTARD Allergen 40 containing 40 mg allergenic substance / kg food sample.|
The SureFood® ALLERGEN Crustaceans PCR has a limit of detection of ≤ 0.4 mg / kg. The limit of quantification is 1 mg / kg using SureFood® PREP Advanced, protocol 1. The limit of detection and limit of quantification were determined using the SureFood® QUANTARD Allergen 40 (matrix: corn flour).
The limit of detection of the complete quantitative determination (DNA extraction and real-time PCR) depends on sample matrix, processing grade, DNA-preparation and DNA-content.