국내기사
넙치 주요 세균성 병원체 4종의 동시 검출을 위한 Multiplex PCR 기법 개발 = Development of multiplex PCR for detecting four bacterial pathogens in olive flounder (Paralichthys olivaceus)
Bacterial diseases pose a major threat to marine aquaculture, causing serious losses in olive flounder (Paralichthys olivaceus). Four major pathogens-Tenacibaculum maritimum, Edwardsiella piscicida, Vibrio harveyi, and Streptococcus parauberis-are associated with severe mortality and production decline. Conventional diagnostic methods such as culture, serological assays, and singleplex PCR are often slow, cross-reactive, and unsuitable for mixed infections. Here, we developed a multiplex PCR (mPCR) assay for simultaneous detection of four pathogens. Species-specific primers were de- signed or adapted, and assay conditions were optimized for clear amplicon separation. The mPCR showed high specificity without cross-reactivity to 15 related bacteria and detected as low as 5.0× 10^1 copies/µL for T. maritimum, and 10^2 copies/µL for the others. It also identified all four pathogens in spiked olive flounder tissues (spleen and kidney) and environmental samples (seawater and biofilm) at 10² cells/g or 10² cells/L. This mPCR provides a rapid, sensitive, and cost-effective diagnostic platform for early detection and on-farm monitoring of bacterial diseases in olive flounder aquaculture.