引用本文:	李秀芹,于志刚,甄毓,蔡青松,米铁柱,李荣秀.应用双特异分子探针技术定性定量检测圆海链藻(Thalassiosira rotula).海洋与湖沼,2009,40(1):12-18.

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*应用双特异分子探针技术定性定量检测圆海链藻(Thalassiosira rotula)*
李秀芹¹, 于志刚², 甄毓², 蔡青松³, 米铁柱⁴, 李荣秀⁵
1.中国海洋大学生命科学与技术学部;2.中国海洋大学化学化工学院;3.中国科学院上海生命科学研究院;4.中国海洋大学环境科学与工程学院;5.上海交通大学生命科学技术学院

摘要:

设计了一套圆海链藻(Thalassiosira rotula)特异性探针, 运用双特异分子探针技术, 对圆海链藻(Thalassiosira rotula)进行了定性定量检测。结果表明,本实验中设计的一套探针与其它十几种藻无交叉反应, 具有种特异性; 细胞裂解液直接杂交检测优于提纯RNA 样品检测; 对自然样品做了初步检测, 发现天然海水中的其它浮游生物对该检测方法影响很小。

关键词: 圆海链藻, 双特异分子探针技术, 赤潮藻检测

DOI：10.11693/hyhz200901003003

分类号:

基金项目:国家高技术研究发展计划(863 计划)资助项目, 2001AA635090 号; 国家高技术研究发展计划(863 计划)资助项目, 2003AA635010 号; 自然科学基金资助项目, 40406028 号。

附件

DETECTION AND QUANTIFICATION OF THALASSIOSIRA ROTULA WITH DOUBLE SPECIFIC PROBE ASSAY

LI Xiu-Qin¹, YU Zhi-Gang², ZHEN Yu², CAI Qing-Song³, MI Tie-Zhu⁴, LI Rong-Xiu⁵

1.Marine Life Sciences College, Ocean University of China;2.Chemistry and Chemical Engineering College, Ocean University of China;3.Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences;4.Environmental Science and Engineering College, Ocean University of China;5.School of Life Science and Biotechnology, Shanghai Jiao Tong University

Abstract:

A double specific probe assay was applied to detect and quantify Thalassiosira rotula, a common species of harmful algae bloom, in which a set of oligonucleotide probes targeting 18S rRNA was used. The probes are species-specific, without obvious incident cross-reactivity with other species. Results of detection on the same number of cells to crude cell lysate and extracted total RNA were compared, showing that the former was better. A preliminary test was made on natural samples, and the noise produced by complex field background was minimal.

Key words: Thalassiosira rotula, Double specific probe assay, Harmful algae detection