| CHEN Jie,ZHEN Yu,MI Tiezhu,YU Zhigang. 2009. Detection of Prorocentrum donghaiense using sandwich hybridization integrated with nuclease protection assay. Acta Oceanologica Sinica, (2):121－126 |
| Detection of Prorocentrum donghaiense using sandwich hybridization integrated with nuclease protection assay |
| Detection of Prorocentrum donghaiense using sandwich hybridization integrated with nuclease protection assay |
| Received:March 25, 2008 Revised:June 16, 2008 |
| DOI： |
| Key words:Prorocentrum donghaiense ribosomal RNA S1 enzyme sandwich hybridization integrated with nuclease protection assay (NPA-SH) |
| 中文关键词: Prorocentrum donghaiense ribosomal RNA S1 enzyme sandwich hybridization integrated with nuclease protection assay (NPA-SH) |
| 基金项目:The National High Technology Research and Development Program ("863" Program) of China under contract No. 2006AA09Z178; the National Natural Science Foundation of China under contract No. 40706044. |
| Author Name | Affiliation | E-mail | | CHEN Jie | College of Marine Life Science, Ocean University of China, Qingdao 266003, China | | | ZHEN Yu | College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China | | | MI Tiezhu | College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China | | | YU Zhigang | College of Chemistry and Chemical Engineering, Ocean University of China, Qingdao 266100, China | zhigangyu@ouc.edu.cn |
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| Abstract: |
| Prorocentrum donghaiense is an important harmful algae bloom (HAB) causing creature in China's seas, and the conventional visual detection can not cope with long-term monitoring and highthroughput sampling projects. An assay for P. donghaiense with sandwich hybridization integrated with nuclease protection assay (NPA-SH) was established. Tests with mixed samples and spiked field ones confirmed its good specificity and sensitivity. The cell number of P. donghaiense correlated well with the optical density, and the regression equation is y=4×10-6x+ 0.694 9, in which x is the cell number, and y is the optical density, with r2=0.953 5. These results show that the NPA-SH method has good feasibility in the detection of P. donghaiense. Results of NPA-SH and microscopy are excellent for each sample. The NPA-SH method was a simple way in quantitative detection of P. donghaiense, and the whole process could be finished in about six hours, which provided a new approach in high-throughput sampling and long-term monitoring of P. donghaiense. |
| 中文摘要: |
| Prorocentrum donghaiense is an important harmful algae bloom (HAB) causing creature in China's seas, and the conventional visual detection can not cope with long-term monitoring and highthroughput sampling projects. An assay for P. donghaiense with sandwich hybridization integrated with nuclease protection assay (NPA-SH) was established. Tests with mixed samples and spiked field ones confirmed its good specificity and sensitivity. The cell number of P. donghaiense correlated well with the optical density, and the regression equation is y=4×10-6x+ 0.694 9, in which x is the cell number, and y is the optical density, with r2=0.953 5. These results show that the NPA-SH method has good feasibility in the detection of P. donghaiense. Results of NPA-SH and microscopy are excellent for each sample. The NPA-SH method was a simple way in quantitative detection of P. donghaiense, and the whole process could be finished in about six hours, which provided a new approach in high-throughput sampling and long-term monitoring of P. donghaiense. |
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