三类分子探针对常见赤潮生物的检测

集中使用寡核苷酸、肽核酸和细胞凝素3类探针对来自东海和厦门海域的现场赤潮样品进行了检测,尝试鉴定识别自然水样中有害的赤潮原因种塔玛亚历山大藻,微小原甲藻和纤小裸甲藻,建立和优化了这些探针的检测方法和样品处理程序.结果表明,在东海和厦门海域的赤潮样品中均成功地检出了塔玛亚历山大藻的分布情况,各探针的检测效率为DBA>Tama28S>Tama5S;在东海和厦门海域的赤潮样品中,也成功地检测出了微小原甲藻,各探针的检测效率为:ConA>PM18S02>PM28S02;在厦门海域的赤潮水样中检出了纤小裸甲藻,各探针的检测效率为:WGA>PNATP28S01>TP18S02>TP28S01.各探针检测结果与相关文献的报道吻合较好.比较这3类探针的特异性,其中以PNA探针为最好,其次为DNA;lectin探针的特异性相对较弱.

Detection of representative harmful algae using three kinds of molecular probes

Molecular probes are useful to separate closely related harmful algal bloom species. Moreover, they can rapidly and clearly differentiate non toxic from toxic algae and to monitor the development of algal blooms in coastal water s. There are three kinds of molecular probes which can be used for identification of HAB species, such as lectin, DNA and peptide nucleotide acid(PNA) probe. These probes are relatively new, and have higher sensitivity, veracity and specificity than traditional taxonomic techniques. In the present study, probes of oligonucleotide, lectin and PNA were applied to detect Prorocentrum minimum, Takayama pulchellum and Alexandrium tamarense of red t ide samples from the East China Sea, Eastern Fujian Coastal Waters and Xiamen Bay, series of studies were undertaken in order to identify and classify these harmful algal species, and the sample processing and detection protocols were evaluated and optimized both in the laboratory and in the field. The results showed that target species of A. tamarense was detected successfully in amples of the East China Sea and Xiamen Bay, and the detection efficiency of different probes specific to A. tamarense was in order: DBA (lectin) > Tama28 S (oligonucleotide probe) > Tama5S(oligonucleotide probe). The target species of P. minimum, was also detected successfully in samples of Eastern Fujian Coastal Waters and Xiamen Bay, and the detection efficiency of different probes specific to P. minimum was as follow: ConA (lectin) > PM18S02 (oligonucleotide) > PM28S02 (oligonucleotide). The target species of T. pulchellum, which was firstly isolated and unialgal cultivated during blooms in Xiamen Bay in 2003 and 2004, was also identified distinctly in samples of Xiamen Bay during blooms in 2005, and the detection efficiency of different probes specific to T. pulchellum was in this order: WGA (lectin) > PNATP28S01 (PNA) > TP18S02 (oligonucleotide)> TP28S01(oligonucleotide). The conclusions were as follow: These target harmful algae bloom species cells appearance in the East China Sea, Eastern Fujian Coastal Waters and Xiamen Bay identified by three probes were correlated well to prevenient results of references, which suggested that specificity of these probes was good and the detecting methods were accurate and reliable. The specificity of three molecular probes was that PNA probe > oligonucleotide probe > lectin probe in order. Thereby, these probes can be used for monitoring marine population dynamics of P. minimum, T. pulchellum and A. tamarense rapidly in future.