Simultaneous Detection of Eight Dairy-Derived Components Using Double-Tube Multiplex qPCR Based TaqMan Probe
Open Access
Abstract
The authentication of milk and dairy products has great significance for food fraud. The present investigation entailed the development of a novel method that amalgamates the double-tube approach with multiplex real-time polymerase chain reaction (PCR) technology, incorporating TaqMan probes, to facilitate the high-throughput screening and detection of animal-derived constituents within milk and dairy products. Eight dairy-derived animal-specific primers and probes were designed for the mitochondrial cytochrome b (Cytb) gene of eight dairy products, including cow, buffalo, yak, goat, sheep, horse, donkey, and camel. Through the developed double-tube detection assays, the above eight targets could be simultaneously identified with a detection limit of 0.00128–0.0064 ng/μL. The multiplex qPCR assay was effectively validated using simulated adulterated samples with different mixing ratios and demonstrated a detection limit of 0.1%. Upon analysis of 54 commercially available dairy products, a mislabeling rate of 33% was revealed. This method affords an efficacious means of detecting dairy product ingredients, thereby offering robust technical backing for market oversight and regulatory enforcement of milk and dairy products.Keywords
Funding Information
- Special National Key Research and Development Plan (2022YFD1600104, 2022A02006-3-1, CARS-36, ASTIP-IAS12, 2022-YWF-ZX-06)
- Xinjiang science and technology projects (2022YFD1600104, 2022A02006-3-1, CARS-36, ASTIP-IAS12, 2022-YWF-ZX-06)
- Earmarked fund for CARS (2022YFD1600104, 2022A02006-3-1, CARS-36, ASTIP-IAS12, 2022-YWF-ZX-06)
- Agricultural Science and Technology Innovation Program (2022YFD1600104, 2022A02006-3-1, CARS-36, ASTIP-IAS12, 2022-YWF-ZX-06)
- Central Public-interest Scientific Institution Basal Research Fund (2022YFD1600104, 2022A02006-3-1, CARS-36, ASTIP-IAS12, 2022-YWF-ZX-06)
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