Development of a quantification assay for the cysts of the toxic dinoflagellate Alexandrium tamarense using real-time polymerase chain reaction

被引:27
|
作者
Kamikawa, R [1 ]
Hosoi-Tanabe, S
Nagai, S
Itakura, S
Sako, Y
机构
[1] Kyoto Univ, Lab Marine Microbiol, Div Appl Biosci, Grad Sch Agr, Kyoto 6068502, Japan
[2] Univ Shiga Prefecture, Sch Environm Sci, Dept Ecosyst Studies, Shiga 5228533, Japan
[3] Res Inst Fisheries & Environm Inland Sea, Harmful Algal Bloom Div, Tox Phytoplankton Sect, Hiroshima 7390452, Japan
关键词
Alexandrium tamarense; cyst; dinoflagellate; paralytic shellfish poisoning (PSP); real-time PCR;
D O I
10.1111/j.1444-2906.2005.01055.x
中图分类号
S9 [水产、渔业];
学科分类号
0908 ;
摘要
The cysts of toxic dinoflagellate Alexandrium tamarense are the seed population for the bloom responsible for paralytic shellfish poisoning (PSP). However, it is impossible to identify the Alexandrium spp. cyst on the basis of morphological features. In this study, we prepared A. tamarense cysts by sexual conjugation in laboratory conditions and developed an efficient DNA extraction method for polymerase chain reaction (PCR) assay. Using the A. tamarense cysts, we established the identification and quantification method showing the species specificity and the high sensistivity for A. tamarense cysts using real-time PCR. This assay was also able to detect and quantify the A. tamarense cysts accurately when mixed with excess cysts of A. catenella (Whedon and Kofoid) Balech prepared by conjugation experiment.
引用
收藏
页码:987 / 991
页数:5
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