Detection of Alexandrium tamarensis by rapid PCR analysis.
Detection of Alexandrium tamarensis by rapid PCR analysis.
复制标题
通过快速 PCR 分析检测塔玛亚历山大藻。
DOI：
10.2144/99261st01
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发表时间：
1999
期刊：
影响因子：
2.7
通讯作者：
T. Murray
中科院分区：
文献类型：
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作者：
S. T. Haley;J. F. Cavender;T. Murray
塔玛亚历山大藻（Alexandiumtamarensis）是一种产藻甲藻，广泛分布于世界各地的沿海沃茨。减轻该物种对健康和经济威胁的一个关键因素是开发一种快速可靠的检测方法。本研究简化了用于分离A.塔玛藻核糖体DNA（rDNA）。继代培养A. tamarensis建立在几个沿海大西洋网站的水样。使用用于培养细胞的商业DNA分离试剂盒方案分离甲藻DNA。用特异于A. tamarensis rDNA经测定，该方法便于检测10（-4）ng/μ L的A。tamarensis DNA此外，该试剂盒使A. tamarensis从水源中分离出来，几乎没有信号衰减。这是一种有价值的快速检测A. tamarensis，甚至在细胞数量大到足以进行形态学鉴定之前。
Alexandrium tamarensis is a toxigenic dinoflagellate found in coastal waters worldwide. A critical factor in alleviating the health and economic threats posed by this species is the development of a rapid and reliable method for detection. This study stream-lined a labor- and resource-intensive protocol for the isolation of A. tamarensis ribosomal DNA (rDNA). Subcultures of A. tamarensis were established in water samples from several coastal Atlantic sites. A commercial DNA isolation kit protocol for cultured cells was used for isolation of the dinoflagellate DNA. Samples were amplified by PCR using primers specific for a 700-bp sequence of A. tamarensis rDNA. It was determined that this method facilitated the detection of 10(-4) ng/microL of A. tamarensis DNA. Furthermore, the kit enabled A. tamarensis to be isolated from the water sources with little signal degradation. This is a valuable technique for the rapid detection of A. tamarensis, even before cell numbers are large enough for morphological identification.