Conventional Detection of Microalgaes Using Nuclease Protection Assay Sandwich Hybridization

Title
Conventional Detection of Microalgaes Using Nuclease Protection Assay Sandwich Hybridization
Author(s)
박미례; 서승석; 황진익; 이택견
KIOST Author(s)
Lee, Taek Kyun(이택견)
Publication Year
2015-09-30
Abstract
The frequent occurrence of harmful algal blooms (HABs) at worldwide has serious damages on the marine environment and fishery resources. A variety of harmful algae including Heterocapsa triquetra, Heterosigma akashiwo, Chattonella marina, Scrippsiella trochoidea have been revealed as causative sources of HABs in Korea’s coasts, but their existence have not been tested in large number of samples and wide scale of marine environment using conventional approaches including light and electron microscopy in long-term monitoring and high-throughput sampling projects. In this study, an integrated sandwich hybridization and nuclease protection assay (NPA-SH) was established for the quantitative detection of H. triqeutra, H. akashiwo, C. marina, S. trochoidea. The specificity of the probes was verified with individual and mixed cultures as well as field collection from Goseong Bay in South Korea. And the quantity of four microalgaes determined by NPA-SH analysis showed such a good correlation with cell-counting by a light microscope. In addition a standard curves for four microalgaes was demonstrated to represent the correlation between optical absorbance in the NPA-SH assay and cell density. These results show that the NPA-SH method is reliable, specific, and precise in the detection of H. triqeutra, H. akashiwo, C. marina, S. trochoidea. Thus, this approach provides an effective alternative to traditional, morphology-basecrippsiella trochoidea have been revealed as causative sources of HABs in Korea’s coasts, but their existence have not been tested in large number of samples and wide scale of marine environment using conventional approaches including light and electron microscopy in long-term monitoring and high-throughput sampling projects. In this study, an integrated sandwich hybridization and nuclease protection assay (NPA-SH) was established for the quantitative detection of H. triqeutra, H. akashiwo, C. marina, S. trochoidea. The specificity of the probes was verified with individual and mixed cultures as well as field collection from Goseong Bay in South Korea. And the quantity of four microalgaes determined by NPA-SH analysis showed such a good correlation with cell-counting by a light microscope. In addition a standard curves for four microalgaes was demonstrated to represent the correlation between optical absorbance in the NPA-SH assay and cell density. These results show that the NPA-SH method is reliable, specific, and precise in the detection of H. triqeutra, H. akashiwo, C. marina, S. trochoidea. Thus, this approach provides an effective alternative to traditional, morphology-base methods for the rapid, precise identification and quantification of harmful algal species and could be used to monitor phytoplankton in field surveys.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/25290
Bibliographic Citation
제 27회 분자세포생물학회 발표 논문집, pp.140, 2015
Publisher
분자세포생물학회
Type
Conference
Language
English
Publisher
분자세포생물학회
Related Researcher
Research Interests

Marine virus,Marine pathogens,해양바이러스,해양병원체

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