Browse > Article
http://dx.doi.org/10.5762/KAIS.2019.20.8.627

Rapid Detection of Cochlodinium polykrikoides by Sandwich Hybridization Integrated with Invertase Assay  

Kang, Mingyeong (Ecological Risk Research Division, Korea Institute of Ocean Science and Technology)
Park, Mirye (Microbial Research Department, Nakdonggang National IOnstitute of Biological Resources)
Lee, Taek-Kyun (Ecological Risk Research Division, Korea Institute of Ocean Science and Technology)
Publication Information
Journal of the Korea Academia-Industrial cooperation Society / v.20, no.8, 2019 , pp. 627-633 More about this Journal
Abstract
Rapid and accurate identification of marine microalgae causing harmful algal blooms (HABs) is a crucial tool for predicting and managing HABs. We previously developed a nuclease protection assay sandwich hybridization (NPA-SH) method for the in situ detection of blooming microalgae Cochlodinium polykrikoides. In this study, we improved the applicability of the NPA-SH method for the detection of C. polykrikoides by simplifying the reaction step. For this purpose, invertase (INV) was conjugated to the signal probe instead of using fluorescence, and sucrose was used as a reactant to induce a color reaction. The INV-signal probe conjugation was confirmed by SDS-PAGE and epifluoromicroscopy. The treatment time and appropriate amounts of the probe and sucrose that optimized the reaction were determined. As a result, the developed INV-SH reduced the treatment time in the field compared with NPA-SH, and also enabled the use of a relatively small volume and low-priced personal glucose meter, as well as an absorbance meter. INV-SH is the first C. polykrikoides species identification technology to which INV has been applied and could be an improved field technique.
Keywords
HABs; C. polykrikoides; NPA-SH; INV-SH; Invertase;
Citations & Related Records
연도 인용수 순위
  • Reference
1 S. Diercks, L. K. Medlin and K. Metfies. "Colorimetric detection of the toxic dinoflagellate Alexandrium minutum using sandwich hybridization in a microtiter plate assay" Harmful Algae, vol. 7, no. 2, pp. 137-145, 2008. DOI: https://doi.org/10.1016/j.hal.2007.06.005   DOI
2 Q. Cai, R. Li, Y. Zhen, T. Mi and Z. Yu. "Detection of two Prorocentrum species using sandwich hybridization integrated with nuclease protection assay" Harmful Algae, vol. 5, no. 3, pp. 300-309, 2006. DOI: https://doi.org/10.1016/j.hal.2005.08.002   DOI
3 Y. Zhen, T. Mi and Z. Yu. "Detection of Phaeocystis globosa using sandwich hybridization integrated with nuclease protection assay (NPA-SH)" J. Environmental Science, vol. 20, no. 12, pp. 1481-1486, 2008. DOI: https://doi.org/10.1016/S1001-0742(08)62553-X   DOI
4 Y. Zhen, T. Mi and Z. Yu. "Detection of several harmful algal species by sandwich hybridization integrated with a nuclease protection assay" Harmful Algae, vol. 8, np. 5, pp. 651-657, 2009. DOI: https://doi.org/10.1016/j.hal.2008.12.001   DOI
5 S. S. Suh, M. Park, J. Hwang, E. J. Kil, S. Lee and T.-K. Lee. "Detection of the dinoflagellate, Cochlodinium polykrikoides, that forms algal blooms using sandwich hybridization integrated with nuclease protection assay" Biotechnology Letters, vol. 38, no. 1, pp. 57-63, 2016. DOI: https://doi.org/10.1007/s10529-015-1947-5   DOI
6 M. Park, S. Y. Park, J. Hwang, S. W. Jung, J. Lee, M. Chang and T.-K. Lee. "Integration of the nuclease protection assay with sandwich hybridization (NPA-SH) for sensitive detection of Heterocapsa triquetra " Acta Oceanologica Sinica, vol. 37, no. 5, pp. 107-112, 2018. DOI: https://doi.org/10.1007/s13131-018-1167-7   DOI
7 Y. Xiang and Y. Lu. "Using personal glucose meters and functional DNA sensors to quantify a variety of analytical targets" Nature Chemistry, vol. 3, pp. 697-703, 2011. doi: https://doi.org/10.1038/nchem.1092.   DOI
8 X. Zhu, H. Xu, R. Lin, G. Yang, Z. Lin and G. Chen. "Sensitive and portable detection of telomerase activity in HeLa cells using the personal glucose meter" Chemical Communications, vol. 50, pp. 7897-7899, 2014. DOI: https://doi.org/10.1039/c4cc03553d   DOI
9 J. Su, J. Xu, Y. Chen, Y. Xiang, R. Yuan and Y. Chai. "Personal glucose sensor for point-of-care early cancer diagnosis" Chemical Communications, vol. 48, pp. 6909-6911, 2012. DOI: https://doi.org/10.1039/c2cc32729e   DOI
10 Y. Xiang and Y. Lu. "An invasive DNA approach toward a general method for portable quantification of metal ions using a personal glucose meter" Chemical Communications, vol. 49, pp. 585-587, 2013. DOI: https://doi.org/10.1039/c2cc37156a   DOI
11 K. S. Park, C. Y. Lee and H. G. Park. "Target DNA induced switches of DNA polymerase activity" Chemical Communications, vol. 51, pp. 9942-9945, 2015. DOI: https://doi.org/10.1039/C5CC02060C   DOI
12 J. Bai, L. Liu, Y. Han, C. Jia and C. Liang. "One-step detection of hexokinase activity using a personal glucose meter" Analytical Methods, vol. 10, pp. 2075-2080, 2018. DOI: https://doi.org/10.1039/c8ay00498f   DOI
13 H. Huang, G. Zhao and W. Dou. "Portable and quantitative point-of-care monitoring of Escherichia coli O157:H7 using a personal glucose meter based on immunochromatographic assay" Biosensors Bioelectronics, vol. 107, pp. 266-271, 2018. DOI: https://doi.org/10.1016/j.bios.2018.02.027   DOI
14 K. Ayers, L. L. Rhodes, J. Tyrrell, M. Gladstone and C. Scholin. "International accreditation of sandwich hybridisation assay format DNA probes for microalgae" New Zealand J. Marine and Freshwater Research, vol. 39, no. 6, pp. 1225-1231, 2005. DOI: https://doi.org/10.1080/00288330.2005.9517388   DOI
15 J. V. Tyrrell, L. B. Connell and C. A. Scholin. "Monitoring for Heterosigma akashiwo using a sandwich hybridization assay" Harmful Algae, vol. 1, no. 2, pp. 205-214, 2002. DOI: https://doi.org/10.1016/S1568-9883(02)00012-4   DOI