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Updated: Jul 23, 2025
Development and Testing of Species-specific Quantitative PCR Assays for Environmental DNA Applications
Published on: November 5, 2020
Development and intralaboratory validation of three Arcidae species using a multiplex polymerase chain reaction assay
Ga-Young Lee1, Seung-Min Yang1, Hae-Yeong Kim1
1Institute of Life Sciences & Resources and Department of Food Science & Biotechnology, Kyung Hee University, Yongin, 17104 Republic of Korea.
Abstract:
Recently, various commercial ark shell products were being sold, and in the case of processed foods, the loss of morphological traits makes species identification visually challenging, which can lead to seafood fraud. Therefore, a multiplex polymerase chain reaction (PCR) assay was developed to simultaneously identify three ark shells. The specific PCR amplicon sizes of the generated species-specific primer pairs were observed to be 99 bp for Anadara kagoshimensis, 148 bp for Anadara broughtonii, and 207 bp for Tegillarca granosa. Specificity was confirmed for 17 fish and shellfish, and only the target was amplified without cross-reactivity. The detection limit for the multiplex PCR assay was 1 pg. Furthermore, 31 commercial products were evaluated to assess the developed assay's applicability. Therefore, the analytical approach used in this study can rapidly and accurately identify ark shells in commercial food, and may be used as an authentication tool in the seafood industry.
Supplementary Information:
The online version contains supplementary material available at 10.1007/s10068-023-01269-2.
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