Matrix-normalised quantification of species by threshold-calibrated competitive real-time PCR: Allergenic peanut in food as one example

摘要: Abstract A novel method to quantify species or DNA on the basis of a competitive quantitative real-time polymerase chain reaction (cqPCR) was developed. Potentially allergenic peanut in food served as one example. Based an internal sequence for normalisation extraction and amplification, cqPCR threshold-calibrated against 100 mg/kg incurred milk chocolate. No external standards were necessary. The molecule successfully calibrator quantification, matrix normalisation, inhibition control. Although designed verification virtual threshold 100 mg/kg, allowed quantification 10–1000 mg/kg various matrices without further adaption: On four PCR replicates per sample, mean recovery chocolate, vanilla ice cream, cookie dough, cookie, muesli 87% (range: 39–147%) comparison 199% 114–237%) by three commercial ELISA kits.