Development of a sensitive and specific qPCR method to detect duck and goose DNA in meat and feathers

Abstract

This study developed a method to identify the species of duck and goose DNA extracted from various matrices. The research material included meat, down, whole feathers, and blood. The analysis was performed with real-time PCR using minor groove binder probes and primers that bind to cytochrome b (duck) and 12S ribosomal RNA (goose) sequences. For all matrices, the reactions were species-specific. Cross-reactions either did not occur or occurred after 36 cycles, corresponding to an amplification for the species content of 10− 5 %, i.e., several times below the limit of detection (LOD = 0.01%) and limit of quantification (LOQ = 0.1%). The standard curves obtained by consecutive dilutions were linear (R2 > 0.99), the slopes ranged from 3.2 to − 3.6, and the retardation factors were 23–26. The amplitude threshold analysis demonstrated that significant differences between successive matrices make it impossible to select one reference material to produce a standard curve effective for analyzing each type of matrix. A comparison between the reference material and the test material reveals that the accuracy of the developed method (the match between the actual and the measured value) is above 78%. Validation methods indicate that this method can be used to identify duck and goose DNA in the analyzed matrices.