Detection of Cochlodinium polykrikoides using sandwich hybridization integrated with nuclease protection assay

Title
Detection of Cochlodinium polykrikoides using sandwich hybridization integrated with nuclease protection assay
Author(s)
서승석; 박미례; 황진익; 김성경; 이택견
KIOST Author(s)
Lee, Taek Kyun(이택견)
Publication Year
2015-09-30
Abstract
The frequent occurrence of harmful algal blooms (HABs) worldwide has serious impacts on the marine environment and fishery resources. Cochlodinium polykrikoides is an important causative agent of HABs in Korea’s seas, but the presence of this organism cannot be tested in large number of samples using conventional approaches including light and electron microscopy in long-term monitoring and high-throughput sampling projects. As such, in this study, an integrated sandwich hybridization and nuclease protection assay (NPA-SH) was established for the quantitative detection of C. polykrikoides. The specificity of the probes was verified with individual and mixed cultures as well as field collection from Gosung Bay, Korea, and the quantity of C. polykrikoides determined by NPA-SH analysis showed a good correlation with that determined by cell-counting with a light microscope. In addition a standard curve for C. polykrikoides was established to represent the correlation between optical absorbance in the NPA-SH assay and cell density. The results show that the NPA-SH method is reliable, specific, and accurate in the detection of C. polykrikoides thus, this approach provides an efficient alternative to traditional, morphology-based methods for the rapid identification and quantification of harmful algal species and could be used to monitor phytoplankton in field surveys.his organism cannot be tested in large number of samples using conventional approaches including light and electron microscopy in long-term monitoring and high-throughput sampling projects. As such, in this study, an integrated sandwich hybridization and nuclease protection assay (NPA-SH) was established for the quantitative detection of C. polykrikoides. The specificity of the probes was verified with individual and mixed cultures as well as field collection from Gosung Bay, Korea, and the quantity of C. polykrikoides determined by NPA-SH analysis showed a good correlation with that determined by cell-counting with a light microscope. In addition a standard curve for C. polykrikoides was established to represent the correlation between optical absorbance in the NPA-SH assay and cell density. The results show that the NPA-SH method is reliable, specific, and accurate in the detection of C. polykrikoides thus, this approach provides an efficient alternative to traditional, morphology-based methods for the rapid 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/25292
Bibliographic Citation
제 27회 분자세포생물학회 발표 논문집, pp.136, 2015
Publisher
분자세포생물학회
Type
Conference
Language
English
Publisher
분자세포생물학회
Related Researcher
Research Interests

Marine virus,Marine pathogens,해양바이러스,해양병원체

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