Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5765694 | Harmful Algae | 2017 | 10 Pages |
Abstract
The aim of this study was to develop a loop-mediated isothermal amplification (LAMP) combined with a chromatographic lateral flow dipstick (LFD) assay to rapidly and specifically detect the Karlodinium veneficum ITS gene. Four groups of LAMP primers were specially designed to target the K. veneficum ITS gene. The LAMP-LFD detection limit was 7.4 pg/μL (approximately 6.5 cells/mL) of K. veneficum genomic DNA and was 10 times more sensitive than standard PCR. The LAMP-LFD method exhibited high specificity and accurately identified K. veneficum algal isolates, but not other algal isolates. To test the assay's accuracy, samples from positive results were further analyzed by sequencing and phylogenetic analysis, all of which were identified as K. veneficum. Over all, the LAMP-LFD assay established in this paper can be used as a reliable and simple method to detect the K. veneficum.
Keywords
POCTITSFITCpractical salinity unitsPSUH7N9LFDFIPBiPInternal transcribed spacerethidium bromideagarose gel electrophoresisPoint-of-care testingLoop mediated isothermal amplificationHarmful algaeLateral Flow DipstickAgefluorescein isothiocyanateLAMPKarlodinium veneficumNeighbour-joiningpolymerase chain reactionPCRavian influenza virus
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Authors
Hai-Long Huang, Peng Zhu, Cheng-Xu Zhou, Shan He, Xiao-Jun Yan,