Development and comparison of a porcine gelatin detection system targeting mitochondrial markers for Halal authentication. (November 2018)

• Record Type: Journal Article
• Authors: Kang, Su San Na; Lee, Hyeon Gyu; Kim, Hyunsook
• Is Part Of: Lebensmittel-Wissenschaft + Technologie =. Volume 97(2018)
• Page Start: 697
• Page End: 702
• Publication Date: 2018-11
• Subjects: Porcine gelatin -- Halal products -- Mitochondrial DNA markers -- D-loop; Food industry and trade -- Periodicals; Food -- Composition -- Periodicals; Microbiology -- Periodicals; Nutrition -- Periodicals; 664.005
• DOI: 10.1016/j.lwt.2018.07.062
• Languages: English
• ISSNs: 0023-6438

Abstract

Abstract: This study aimed to compare the sensitivity of porcine-specific primers targeting different mitochondrial DNA for the detection of porcine gelatin in ramen stock powder designated "Halal" using SYBR Green I real-time PCR. Four mitochondrial DNA markers (D-loop, ATP8, ND5, and 12S rRNA) were used to detect the porcine gelatin DNA. Extraction of DNA from porcine gelatin and spiked mixtures was successfully performed using a DNeasy Mericon Food Kit. Each primer had high porcine specificity for D-loop, ATP8, and ND5 genes, but not the 12S rRNA gene. Also, the detection limit for each primer was tested in a serial dilution of porcine gelatin DNA (10, 1, 0.1, 0.05, 0.01, and 0.005 ng) and ramen stock powder spiked with porcine gelatin (1, 0.1, 0.05, and 0.025%). Assays targeting both D-loop and ND5 genes were able to detect a level as low as 0.01 ng, while the detection limit for the ATP8 gene was 0.05 ng in pure porcine gelatin. The detection limits of D-loop, ATP8, and ND5 genes were 0.05%, 0.1%, and 1% (w/w) in ramen stock powder spiked with porcine gelatin, respectively. In conclusion, a novel detection system targeting the mitochondrial D-loop was superior to the others and could be an effective and simple method to authenticate Halal products. Highlights: A porcine gelatin detection system was developed based on mitochondrial genes. Among four assays, mitochondrial D-loop assay was the most reliable and sensitive. This assay will provide simple and efficient detection system. This assay will authenticate halal products including ramen stock powder.