Improved triplex real‐time PCR with endogenous control for synchronous identification of DNA from chicken, duck, and goose meat

Abstract

The authentication and labeling of meat products, concerning origins and species, are key to fair trade and to protect consumer interests in the market. We developed an improved triplex real‐time PCR approach to simultaneously identify chicken, duck, and goose DNA in meat, including an endogenous control to avoid false negatives. Our method specifically detected DNA from chicken, duck, and goose, and showed no cross‐reaction with DNA extracted from other meat types. The detection limits of chicken, duck, and goose DNA were 0.001–0.00025 ng, 0.0025–0.0001 ng, and 0.001–0.00001 ng, respectively, and we were able to simultaneously identify DNA from two distinct origins using as little as 0.1% of total meat weight. Our newly generated triplex real‐time PCR method with endogenous control exhibited high specificity, sensitivity, and efficiency for simultaneous identification of DNA from chicken, duck, and goose in meat.

Molecular Authentication of Twelve Meat Species Through a Promising Two-Tube Hexaplex Polymerase Chain Reaction Technique

“…Polymerase chain reaction (PCR) techniques such as species-specific PCR, multiplex PCR, PCR-RFLP, real-time PCR, random amplified polymorphic DNA (RAPD), and DNA barcoding have evolved as preferred methods for meat fraud detection ( 2 , 10 , 11 ). Recently, both multiplex and real-time PCR techniques have been widely applied for meat fraud detection ( 12 ). Real-time PCR provides more detailed information regarding the identification and quantification of meat species ( 13 ).…”

Design and development of a rapid meat detection system based on RPA-CRISPR/Cas12a-LFD

“…At the present stage, nucleic acid-based detection methods for meat are developing rapidly, with routine PCR assays for chicken, duck, pork, beef and lamb ( Ulca et al, 2013 ; Dai et al, 2015 ), but direct PCR methods require complex manipulation and gel electrophoresis, which is time-consuming and labor-intensive. Multiplex-PCR( Uddin et al, 2021 ; Cheng et al, 2022 ), real-time fluorescence quantitative PCR ( Liu et al, 2021 ; Li et al, 2021 ) and droplet digital PCR ( Hu et al, 2021 ; Shehata et al, 2017 ) which produce higher specificity than direct PCR, have also been well developed in meat adulteration. Compared to direct PCR, the accuracy, sensitivity and detection efficiency of the later derived PCR techniques in meat adulteration detection are significantly enhanced.…”

Rapid detection of duck ingredient in adulterated foods by isothermal recombinase polymerase amplification assays

“…The exo probe for real-time RPA and the nfo probe for LFS RPA were designed based on the amplified region of the optimal primer pairs. Besides the aforementioned primers and probes, the real-time PCR primers and TaqMan probe from the previously described protocol were also synthesized in this study ( Liu et al, 2021 ). The related nucleotide sequences are shown in Table 2 .…”