A rapid real-time PCR method to differentiate between mottled skate (Beringraja pulchra) and other skate and ray species

Mi-Ra Kim; Kisung Kwon; Yoo-Kyung Jung; Tae Sun Kang

doi:10.1016/j.foodchem.2018.02.056

A rapid real-time PCR method to differentiate between mottled skate (Beringraja pulchra) and other skate and ray species

Food Chem. 2018 Jul 30:255:112-119. doi: 10.1016/j.foodchem.2018.02.056. Epub 2018 Feb 12.

Authors

Mi-Ra Kim¹, Kisung Kwon¹, Yoo-Kyung Jung¹, Tae Sun Kang²

Affiliations

¹ New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety, 187 Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do 28159, South Korea.
² New Hazardous Substance Team, Food Safety Evaluation Department, National Institute of Food and Drug Safety Evaluation, Ministry of Food and Drug Safety, 187 Osongsaengmyeong 2-ro, Osong-eup, Heungdeok-gu, Cheongju-si, Chungcheongbuk-do 28159, South Korea. Electronic address: missa@korea.kr.

PMID: 29571456
DOI: 10.1016/j.foodchem.2018.02.056

Abstract

Skates and rays are commercially important fish in South Korea, and among them, Beringraja pulchra has the highest economic value. However, the similar morphological traits among skates and rays are often exploited for seafood fraud. Here, we designed both Beringraja pulchra-specific and skate-universal primer sets, capable of detecting short sequences in the cytochrome oxidase subunit I gene, and developed highly sensitive and reliable quantitative real-time PCR (qPCR) assays to differentiate between Beringraja pulchra and other skate and ray species. AΔCq method based on differences in the amplification efficiency was developed, validated, and then used to confirm the presence of Beringraja pulchra in twenty-six commercial skate products. The averageΔCq value obtained for other skate species (18.94 ± 3.46) was significantly higher than that of Beringraja pulchra (1.18 ± 0.15). For on-site applications, we developed an ultra-fast qPCR assay, allowing for completion of the entire analytical procedure within 30 min.

Keywords: Amplification efficiency; Cytochrome oxidase subunit I; Genetic identification; Seafood authentication; Ultra-fast qPCR.

MeSH terms

Animals
DNA Primers
Electron Transport Complex IV / genetics
Fish Products / analysis*
Fish Proteins / genetics
Food Analysis / methods*
Real-Time Polymerase Chain Reaction / methods*
Reproducibility of Results
Republic of Korea
Skates, Fish / genetics*
Species Specificity

Substances

DNA Primers
Fish Proteins
Electron Transport Complex IV