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Detection of two diatoms using sandwich hybridization integrated with nuclease protection assay (NPA-SH)

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Abstract

Nuclease protection assay (NPA) probes were designed to target the rRNA of Chaetoceros curvisetus and Skeletonema costatum, and quantitative sandwich hybridization integrated with nuclease protection assay (NPA-SH) was developed to detect C. curvisetus and S. costatum in culture and field samples in Jiaozhou Bay, China. The specificity and validity of the NPA-SH technique were tested with cultured pure strains, mixed strains and field samples, and by comparison with that of microscopy observation. The linear detection range for C. curvisetus was 4.2 × 104 to 1.2 × 106 cells with a detection limit of 42 cells ml−1. The linear range for S. costatum was 6.0 × 104 to 1.0 × 106 cells with a detection limit of 60 cells ml−1. The NPA-SH in this study provides a convenient tool for rapid assessment of HAB species in marine environments.

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Acknowledgements

We thank Dr. Dongyan Liu, Miss Shanying He, Miss Liyuan Zhao and Miss Jing Sun for technical help in this work. We would also like to thank the reviewers for greatly improving this manuscript and their time, patience and valuable feedback. This study was supported by the National High Technology Research and Development Program of China (863 Program, 2001AA635090).

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Authors and Affiliations

  1. College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao, 266003, PR China

    Yu Zhen & Zhigang Yu

  2. School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shangha, 200240, PR China

    Qingsong Cai & Rongxiu Li

  3. College of Environmental Science and Engineering, Ocean University of China, Qingdao, 266003, PR China

    Tiezhu Mi

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  1. Yu Zhen
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  2. Zhigang Yu
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  3. Qingsong Cai
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  4. Tiezhu Mi
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Correspondence to Zhigang Yu.

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Handling editor: D. Hamilton

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Zhen, Y., Yu, Z., Cai, Q. et al. Detection of two diatoms using sandwich hybridization integrated with nuclease protection assay (NPA-SH). Hydrobiologia 575, 1–11 (2007). https://doi.org/10.1007/s10750-006-0278-x

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  • DOI: https://doi.org/10.1007/s10750-006-0278-x

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