An integrated digital polymerase chain reaction chip for multiplexed meat adulteration detection

Meat adulteration detection is a common concern of consumers. Here, we proposed a multiplex digital polymerase chain reaction method and a low-cost device for meat adulteration detection. Using a polydimethylsiloxane microfluidic device, polymerase chain reaction reagents could be pump-free loaded into microchambers (40 x 40 chambers) automatically. Due to the independence of multiplex fluorescence channels, deoxyribonucleic acid templates extracted from different animal species could be distinguished by one test. In this paper, we designed primers and probes for four types of meat (beef, chicken, pork, and duck) and labeled each of the four fluorescent markers (hexachlorocyclohexane [HEX], 6-carboxyfluorescein [FAM], X-rhodamine [ROX], and cyanine dyes 5 [CY5]) on the probes. Specific detection and mixed detection experiments were performed on four types of meat, realizing a limit of detection of 3 copies/mu L. A mixture of four different species can be detected by four independent fluorescence channels. The quantitative capability of this method is found to meet the requirements of meat adulteration detections. This method has great potential for point-of-care testing together with portable microscopy equipment.

Automated summary

Meat adulteration detection is achieved using a multiplex digital polymerase chain reaction method and a low-cost device. A polydimethylsiloxane microfluidic device is used to automatically load polymerase chain reaction reagents into microchambers. The method can distinguish DNA templates from different animal species and has a limit of detection of 3 copies/μL. The method has potential for point-of-care testing with portable microscopy equipment.