| 摘要: |
| 利用本实验室首创的双特异分子探针技术对胶州湾常见的3 种硅藻-旋链角毛藻 (Chaetoceros curvisetus Cleve)、尖刺拟菱形藻 (Pseudo-nitzschia pungens (Grunow ex Cleve) Halse) 和中肋骨条藻 (Skeletonema costatum (Greville) Cleve) 进行定性与定量分析。结果表明, 无论是对实验室样品还是自然样品进行检测, 本技术均有较好的适用性, 对3 种目标藻的最低检测细胞数分别为4.2 × 104 、9.4 × 103 和6.0 × 104 个细胞。统计分析表明, 双特异分子探针技术对3 种目标藻的分析结果与经典的显微镜镜检结果没有显著性差异。本技术利用了多孔酶标板进行检测, 整个实验过程约7 h,可在较短时间内分析大量样品中的多种微藻, 为同时监测多种赤潮藻开辟了一条新途径。
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| 关键词: 胶州湾, 旋链角毛藻, 尖刺拟菱形藻, 中肋骨条藻, 双特异分子探针技术 |
| DOI：10.11693/hyhz201001003003 |
| 分类号: |
| 基金项目:国家十五863 计划资源与环境领域海洋监测主题资助项目, 2001AA635090 号和国家自然科学基金资助项目, 40406028 号。 |
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| DETECTION OF THREE DIATOM SPECIES IN JIAOZHOU BAY USING SANDWICH HYBRIDIZATION INTEGRATED WITH NUCLEASE PROTECTION ASSAY |
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ZHEN Yu1, YU Zhi-Gang2, CAI Qing-Song3, MI Tie-Zhu1, LI Rong-Xiu3
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1.Key Laboratory of Marine Environment and Ecology, Ministry of Education, Ocean University of China;2.Key Laboratory of Marine Chemistry Theory and Technology, Ministry of Education, Ocean University of China;3.School of Life Science and Biotechnology, Shanghai Jiao Tong University
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| Abstract: |
| Sandwich hybridization integrated with nuclease protection assay (NPA-SH) was developed and successfully applied in simultaneously detecting three diatoms species in Jiaozhou Bay [Chaetoceros curvisetus Cleve, Pseudo-nitzschia pungens (Grunow ex Cleve) Halse and Skeletonema costatum (Greville) Cleve] qualitatively and quantitatively. The new method can be well applied in analyzing samples in both laboratory and field. The detection limit to the three species was 4.2×104, 9.4×103 and 6.0×104 cells, respectively. No distinct difference in result between the NPA-SH and the traditional microscopy. The NPA-SH can be completed in a short time of 7 h, in which 96-microplates are used. Therefore, a large amount of samples can be processed for multiple algae, which favors the timely monitoring on harmful algal blooms in marine environment.
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| Key words: Jiaozhou Bay, Chaetoceros curvisetus, Pseudo-nitzschia pungens, Skeletonema costatum, Sandwich Hybridization integrated with Nuclease Protection Assay (NPA-SH) |
