The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Karlodinium veneficum.

Journal: Harmful algae
Published Date: Dec 21, 2016

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

  • Hai-Long Huang
    Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: baiduhailong@sina.com.
  • Peng Zhu
    Center for Drug Safety Evaluation and Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, 201203, China.
  • Cheng-Xu Zhou
    Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: zhouchengxu@nbu.edu.cn.
  • Shan He
    Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: heshan@nbu.edu.cn.
  • Xiao-Jun Yan
    Key Laboratory of Applied Marine Biotechnology, Ningbo University, Ningbo 315211, China. Electronic address: yanxiaojun@nbu.edu.cn.

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