Department of Biology, University of Oslo, P.O. Box 1066, Blindern, 0316, Oslo, Norway.
Environ Sci Pollut Res Int. 2013 Oct;20(10):6778-93. doi: 10.1007/s11356-012-1295-0. Epub 2013 May 11.
Rapid and reliable detection of harmful algae in coastal areas and shellfish farms is an important requirement of monitoring programmes. Monitoring of toxic algae by means of traditional methods, i.e., light microscopy, can be time consuming when many samples have to be routinely analysed. Reliable species identification requires expensive equipment and trained personnel to carry out the analyses. However, all techniques for the monitoring of harmful algae usually require transportation of samples to specialised laboratories. In many monitoring laboratories, results are usually obtained within five working days after receiving the sample and therefore preventative measures are not always possible. Molecular technologies are rapidly improving the detection of phytoplankton and their toxins and the speed at which the results can be obtained. Assays are based on the discrimination of the genetic differences of the different species and species-specific probes can be designed. Such probes have been adapted to a microarray or phylochip format and assessed in several EU monitoring sites. Microarray results are presented for 1 year of field samples validated with cell counts from concentrated samples taken during toxic events from the weekly sampling of the Galician Monitoring Programme done by INTECMAR. The Galician monitoring laboratory does their own counting and their results are posted on their web site within 24 h. There was good correlation between cells present and microarray signals. In the few cases of false negatives, these can be attributed to poor RNA extraction of the target species, viz. Prorocentrum or Dinophysis. Where potential false positives were encountered, the smaller volume taken for cell counts as compared to the upto 300 times more volume taken for RNA extraction for the microarray is likely the cause for these differences, making the microarray more sensitive. The microarray was able to provide better species resolution in Alexandrium and Pseudo-nitzschia. In all cases, the toxins recovered by the toxin array were matched by target species in the array or in the cell counts.
快速可靠地检测沿海地区和贝类养殖场的有害藻类是监测计划的重要要求。通过传统方法（即，光学显微镜）监测有毒藻类，当需要常规分析大量样本时，可能会很耗时。可靠的物种鉴定需要昂贵的设备和经过培训的人员来进行分析。然而，有害藻类监测的所有技术通常都需要将样品运送到专门的实验室。在许多监测实验室中，通常在收到样品后的五个工作日内才能获得结果，因此并非总是可以采取预防措施。分子技术正在快速提高浮游植物及其毒素的检测速度和获得结果的速度。测定基于不同物种遗传差异的区分，并且可以设计物种特异性探针。这些探针已被改编为微阵列或 phylochip 格式，并在几个欧盟监测站点进行了评估。微阵列结果显示了 1 年的野外样本，这些样本是通过 INTECMAR 进行的加利西亚监测计划的每周采样中浓缩样本的细胞计数进行验证的。加利西亚监测实验室自行进行计数，并在 24 小时内将结果发布在其网站上。存在的细胞与微阵列信号之间存在良好的相关性。在少数假阴性的情况下，这些可以归因于目标物种（即，夜光藻或赤潮藻）的 RNA 提取不佳。在遇到潜在的假阳性的情况下，与用于微阵列的多达 300 倍更多体积的 RNA 提取相比，细胞计数所取的较小体积可能是造成这些差异的原因，从而使微阵列更敏感。微阵列能够在亚历山大藻和拟菱形藻中提供更好的物种分辨率。在所有情况下，毒素阵列回收的毒素都与阵列或细胞计数中的目标物种匹配。
