近海赤潮藻全自动分析工作站的研制

本文介绍了以伞新的分析技术--双特异分子探针技术为基础的近海赤潮藻全自动分析工作站.该工作站是将生物技术、机械设计、计算机控制技术、检测技术等有机融合为一体的船载型机电一体化系统.该工作站采用三自由度机械手作为传输机构,集成了海水采样预处理系统及生化分析所需的仪器模块,实现了赤潮藻采样、处理、分析的全自动化.与现有分析手段比较,采用本工作站可大大提高榆测的实时性、准确性及产量,减轻了检测的劳动强度和由于手工重复性劳动带来的人为误差,现已经成功运行.     

运用荧光定量PCR技术检测中肋骨条藻的研究

以rDNA序列为寻找种特异性引物的靶区域,通过分析中肋骨条藻rDNA序列,设计出7套适合用于实时荧光定量PCR方法(RFQ-PCR)的引物与探针,经引物验证,选择Primer6(F/R)进一步分析,对比扩增序列及核酸杂交验证,表明该探针可作为中肋骨条藻特异性探针.最后以Primer6(F/R)和TaqMan6建立了定量检测中肋骨条藻的RFQ-PCR方法,并绘制出了定量检测中肋骨条藻的标准曲线,测定数据用镜检计数进行了验证,证明该方法是准确可靠的.     

Development of real-time PCR method for rapid detection and quantification of Heterosigma akashiwo

To rapidly detect the harmful algae H. akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H. akashiwo were used for designing species-specific primers, and a RFQ-PCR （Realtime Fluorescent Quantitative Polymerase Chain Reaction） method was developed for quantitative detection of H. akashiwo. Primer H. akashiwo and TaqMan probe were designed, and the specificity of primer was checked with PCR. A calibration curve was constructed with cycle threshold value against visual counted cell number. And the value of the curve was tested with other H. akashiwo samples, which were assayed with both the RFQ- PCR method and visual count under microscope.