运用双特异分子探针技术对胶州湾三种硅藻的检测

DETECTION OF THREE DIATOM SPECIES IN JIAOZHOU BAY USING SANDWICH HYBRIDIZATION INTEGRATED WITH NUCLEASE PROTECTION ASSAY

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(Chaeto-ceros curvisetus Cleve)、尖刺拟菱形藻(Pseudo-nitzschia pungens(Grunowex 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7h,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。

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 4.2×10^4, 9.4×10^3 and 6.0×10^4 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.