Development and evaluation of a DNA microarray assay for the simultaneous detection of nine harmful algal species in ship ballast and seaport waters

作者：Chen Xianfeng; Zhou Qianjin; Duan Weijun; Zhou Chengxu; Duan Lijun; Zhang Huili; Sun Aili; Yan Xiaojun; Chen Jiong^*

来源：Chinese Journal of Oceanology and Limnology, 2016, 34(1): 86-101.

DOI：10.1007/s00343-015-4265-9

• ballast waters
• DNA microarray
• harmful algae
• limit of detection
• multiplex PCR
• seaport waters

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 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.

• 出版日期2016-1
• 单位宁波检验检疫科学技术研究院; 宁波大学