摘要
Rapid,high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae,which are responsible for algal blooms,such as red and green tides. In this study,we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously,namely A lexandrium tamarense,Gyrodinium instriatum,Heterosigma akashiwo,Karenia mikimotoi,Prorocentrum donghaiense,Prorocentrum minimum,Ulva compressa,Ulva ohnoi and Ulva prolifera. This method achieved a limit of detection(LOD) of 0.5 ng of genomic DNA(orders of magnitude of the deci-nanogram range) in the tested algae cultures. Altogether,230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%,respectively,relative to conventional morphological methods. This indicated that this high-throughput,automatic,and specific method is well suited for the detection of algae in water samples.
Rapid, high-throughput and reliable methods are urgently required to accurately detect and monitor harmful algae, which are responsible for algal blooms, such as red and green tides. In this study, we successfully developed a multiplex PCR-based DNA microarray method capable of detecting nine harmful algal species simultaneously, namely Alexandrium tamarense, Gyrodinium instriatum, Heterosigma akashiwo, Karenia mikimotoi, Prorocentrum donghaiense, Prorocentrum minimum, Ulva compressa, Ulva ohnoi and Ulvaprolifera. This method achieved a limit of detection （LOD） of 0.5 ng ofgenomic DNA （orders of magnitude of the deci-nanogram range） in the tested algae cultures. Altogether, 230 field samples from ship ballast waters and seaport waters were used to evaluate the DNA microarray. The clinical sensitivity and specificity of the DNA microarray assay in detecting field samples were 96.4% and 90.9%, respectively, relative to conventional morphological methods. This indicated that this high-throughput, automatic, and specific method is well suited for the detection of algae in water samples.
基金
Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA092001)
the Science and Technology Project of Zhejiang Province(No.2013C03045-1)
the Zhejiang Marine Biotechnology Innovation Team(No.2010R50029-12)
the Natural Science Foundation of Ningbo City of China(No.2013A610168)
the KC Wong Magna Fund in Ningbo University
