Development of a quantitative PCR primers and probe for environmental DNA detection of Pacific herring Clupea pallasii

Abstract

An eDNA assay for the detection of Pacific herring (Clupea pallasii) in water samples was developed using quantitative PCR (qPCR) technology. Species-specific primers and a minor groove binding (MGB) probe were designed based on the mitochondrial cytochrome b gene sequence. Upon conducting a cross-species test, the target species DNA was not only detected at low concentrations but also with high specificity. The assay was validated using field-collected water samples from aquatic environments known to be inhabited by Pacific herring and tested positive in samples from Pacific herring-inhabited sites. Our in silico, in vitro, and in situ results highlight the sensitivity of our proposed eDNA-based procedure for the detection of C. pallasii DNA, and could provide a non-invasive and non-destructive approach to detect this species and characterize its distribution in the Jinhaeman Bay, Korea.