Development and comparison of a porcine gelatin detection system targeting mitochondrial markers for Halal authentication

Su San Na Kang, Hyeon Gyu Lee, Hyunsook Kim

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish
Pages (from-to)697-702
Number of pages6
JournalLWT
Volume97
DOIs
StatePublished - 2018 Nov 1

Fingerprint

Gelatin
gelatin
Swine
swine
Powders
Limit of Detection
powders
detection limit
Mitochondrial DNA
genes
Genes
DNA
mitochondrial DNA
ribosomal RNA
Genetic Markers
rRNA Genes
Real-Time Polymerase Chain Reaction
quantitative polymerase chain reaction
Food
genetic markers

Keywords

  • D-loop
  • Halal products
  • Mitochondrial DNA markers
  • Porcine gelatin

Cite this

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title = "Development and comparison of a porcine gelatin detection system targeting mitochondrial markers for Halal authentication",
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.",
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Development and comparison of a porcine gelatin detection system targeting mitochondrial markers for Halal authentication. / Kang, Su San Na; Lee, Hyeon Gyu; Kim, Hyunsook.

In: LWT, Vol. 97, 01.11.2018, p. 697-702.

Research output: Contribution to journalArticle

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