title: The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Karlodinium veneficum.
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 7.4pg/μL (approximately 6.5cells/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.
authors.affiliation: Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: baiduhailong@sina.com.
authors.forename: Hai-Long
authors.initials: HL
authors.lastname: Huang
authors.affiliation: Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: zhupeng@nbu.edu.cn.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: zhouchengxu@nbu.edu.cn.
authors.forename: Cheng-Xu
authors.initials: CX
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: heshan@nbu.edu.cn.
authors.forename: Shan
authors.initials: S
authors.lastname: He
authors.affiliation: Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: yanxiaojun@nbu.edu.cn.
authors.forename: Xiao-Jun
authors.initials: XJ
authors.lastname: Yan
mesh_terms: D002681:China; D002845:Chromatography / Q000295:instrumentation / Q000379:methods*; D021903:DNA, Ribosomal Spacer; D004141:Dinoflagellida / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010802:Phylogeny; D048030:RNA, Algal; D016053:RNA, Protozoan; D012680:Sensitivity and Specificity
chemical_list: D021903:DNA, Ribosomal Spacer; D048030:RNA, Algal; D016053:RNA, Protozoan
keywords: Harmful algae; KV70 primers; Karlodinium veneficum; LAMP-LFD assay