Detection of the dinoflagellate, Cochlodinium polykrikoides, that forms algal blooms using sandwich hybridization integrated with nuclease protection assay.
OBJECTIVE
To detect Cochlodinum polykrikoides in long-term monitoring and high-throughput sampling projects using an integrated sandwich hybridization and nuclease protection assay (NPA-SH).


RESULTS
The specificity of the probes was verified with individual and mixed cultures as well as field collection, 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.


CONCLUSIONS
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.
South Sea Environment Research Department, Korea Institute of Ocean Science & Technology, Geoje, 656-830, Korea.
Sung-Suk
SS
Suh
South Sea Environment Research Department, Korea Institute of Ocean Science & Technology, Geoje, 656-830, Korea.|Marine Environmental Chemistry and Biology, Korea University of Science and Technology, Daejeon, 305-350, Korea.
Mirye
M
Park
South Sea Environment Research Department, Korea Institute of Ocean Science & Technology, Geoje, 656-830, Korea.|Marine Environmental Chemistry and Biology, Korea University of Science and Technology, Daejeon, 305-350, Korea.
Jinik
J
Hwang
Department of Genetic Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea.
Eui-Joon
EJ
Kil
Department of Genetic Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea.
Sukchan
S
Lee
South Sea Environment Research Department, Korea Institute of Ocean Science & Technology, Geoje, 656-830, Korea. tklee@kiost.ac.|Marine Environmental Chemistry and Biology, Korea University of Science and Technology, Daejeon, 305-350, Korea. tklee@kiost.ac.
Taek-Kyun
TK
Lee
D004141:Dinoflagellida / Q000235:genetics / Q000302:isolation & purification*; D005068:Eutrophication; D006824:Hybridization, Genetic; D023282:Nuclease Protection Assays / Q000379:methods*; D016053:RNA, Protozoan / Q000032:analysis
D016053:RNA, Protozoan
Algal blooms; Cochlodinium polykrikoides; Microalgae detection; Ribosomal RNA; S1 enzyme; Sandwich hybridization integrated with nuclease protection assay (NPA-SH)