AUTHOR=Noh Eun Soo , Dong Chun Mae , Park Hee Jeong , Kim Eun Mi , Jung Hyo Sun , Kong Hee Jeong , Kim Young Ok TITLE=Development of species identification techniques for anguillid eels using species-specific genetic markers JOURNAL=Frontiers in Marine Science VOLUME=Volume 12 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/marine-science/articles/10.3389/fmars.2025.1518562 DOI=10.3389/fmars.2025.1518562 ISSN=2296-7745 ABSTRACT=The decline in eel resources, coupled with the challenges of morphological identification and the rise of illegal trade, highlights the urgent need for accurate species identification techniques. To address this, a multiplex PCR assay was developed, targeting the mitochondrial Cytochrome b gene. Species-specific primers were designed and their efficacy validated through single PCR. The multiplex PCR conditions were then optimized to enable the simultaneous amplification of five major eel species. This assay exhibited high accuracy, specificity, and sensitivity, successfully identifying all five species even when DNA concentrations were low. This multiplex PCR assay offers a rapid and cost-effective solution for eel species identification, with the potential to significantly bolster eel conservation efforts. By enabling accurate species identification, it can help combat illegal trade and support the sustainable management of eel resources.