Development of fast and sensitive real-time qPCR assay based on a novel probe for detection of porcine DNA in food sample

- We report a qPCR assay using novel ZEN™ probe to detect porcine adulteration in food.
- The assay was highly specific to porcine DNA.
- Porcine DNA was successfully detected in real pork food samples.
- LOD in standard sample was as low as 1 pg/μl and in the raw pork-chicken binary mixture was as low as 0.001%.
- The assay rapidly detected 10 ng/μl of porcine DNA in approximately 11.33 min.

Porcine adulteration of food is objectionable to a sizeable percentage of the global population owing to health concerns and/or religious faiths. However, unintentional or intentional porcine adulteration is common in the food industry - for which a strong demand prevails for a fast and sensitive method to detect and quantify porcine DNA in foods. In this study, we are reporting the development of a real-time qPCR assay based on the novel ZEN™ probe for the fast and sensitive detection of porcine DNA in real food samples. The assay's specificity to porcine DNA was confirmed against nine species. Standard curve was developed and sensitivity of the assay was tested with six ten-fold dilutions of the DNA standard. Our novel assay successfully detected as low as 1 pg/μl of porcine DNA and as low as 0.001% pork in the raw pork-chicken binary mixture.