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Detection and quantification of cultured marine Alexandrium species by real-time PCR

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Abstract

The occurrence of harmful algal blooms (HABs) throughout the world has increased and poses a large threat to human health, fishery resources and tourism industries. The genus Alexandrium includes a number of toxic species associated with HABs. Therefore, it is very important to rapidly detect and monitor the harmful algae, such as Alexandrium genus. In this study, a standard curve of plasmid containing 18S rDNA-28S rDNA region from Alexandrium catenella was constructed and 5.8S rDNA sequence served as the primer of the real-time PCR. Cultured A. catenella, Alexandrium affine, Alexandrium lusitanicum and Alexandrium minutum samples were analyzed by real-time PCR using the same set of primers simultaneously. Using microscopy cells counts, 5.8S rDNA copies per cell and total DNA per cell were estimated. This assay method is promising for rapid detection of large number of Alexandrium samples.

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Acknowledgments

This research was supported by fund from Key Laboratory of Integrated Marine Monitoring and Applied Technologies for Harmful Algal Blooms,S.O.A., MATHAB (No. MH200804). The authors thank Dr. Yuping Miao for providing Alexandrium minutum AMTK4 and Dr. Tinglin Yang for providing Alexandrium catenella ACDH 01, Alexandrium affine CCMP112 and Alexandrium lusitanicum CCMP1888.

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  1. State Key Laboratory of Microbial Metabolism, Marine Biotechnology Laboratory, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China

    Fengli Zhang & Zhiyong Li

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  1. Fengli Zhang
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  2. Zhiyong Li
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Correspondence to Fengli Zhang.

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Zhang, F., Li, Z. Detection and quantification of cultured marine Alexandrium species by real-time PCR. World J Microbiol Biotechnol 28, 3255–3260 (2012). https://doi.org/10.1007/s11274-012-1136-9

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