Rapid and sensitive detection method for Karlodinium veneficum by recombinase polymerase amplification coupled with lateral flow dipstick.

摘要: Abstract The dinoflagellate Karlodinium veneficum that is usually present at relatively low cell abundances a globally-distributed harmful algal bloom-forming species, which negatively affects marine ecosystems, fisheries, and human health. Hence, an efficient detection platform for the rapid sensitive identification of K. highly demanded. In this study, method referred to as recombinase polymerase amplification coupled with lateral flow dipstick (RPA-LFD) was developed veneficum. primers RPA probe LFD were designed specially target internal transcribed spacer by molecular cloning multiple alignments related sequences. can gain approximately 300 bp specific band from Successful could be achieved temperature range 35 °C–45 °C. 30 min produce enough products generate clearly visible electrophoresis bands adequate subsequent analysis. visually detected naked eyes through after automatic chromatography 5 min room temperature. RPA-LFD exclusively displayed no cross-reactivity other species are commonly distributed along Chinese coast. addition, lowest limit 10 ng μL−1 genomic DNA 0.1 mL−1, 100-fold than conventional PCR. conclusion, assay in study used monitor future.