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Seasonal dynamics of harmful algae in outer Oslofjorden monitored by microarray, qPCR, and microscopy

机译:通过微阵列,qPCR和显微术监测在奥斯陆峡湾外部有害藻类的季节性动态

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Monitoring of marine microalgae is important to predict and manage harmful algal blooms. Microarray Detection of Toxic ALgae (MIDTAL) is an FP7-funded EU project aiming to establish a multi-species microarray as a tool to aid monitoring agencies. We tested the suitability of different prototype versions of the MIDTAL microarray for the monthly monitoring of a sampling station in outer Oslofjorden during a 1-year period. Microarray data from two different versions of the MIDTAL chip were compared to results from cell counts (several species) and quantitative real-time PCR (qPCR; only Pseudochattonella spp.). While results from generation 2.5 microarrays exhibited a high number of false positive signals, generation 3.3 microarray data generally correlated with microscopy and qPCR data, Responsible editor: Philippe Garrigues with three important limitations: (1) Pseudo-nitzschia cells were not reliably detected, possibly because cells were not sufficiently retained during filtration or lysed during the extraction, and because of low sensitivity of the probes; (2) in the case of samples with high concentrations of non-target species, the sensitivity of the arrays was decreased; (3) one occurrence of Alexandrium pseiidogonyanlax was not detected due to a 1-bp mismatch with the genus probe represented on the microarray. In spite of these shortcomings our data demonstrate the overall progress made and the potential of the MIDTAL array. The case of Pseudochattonella - where two morphologically similar species impossible to separate by light microscopy were distinguished - in particular, underlines the added value of molecular methods such as microarrays in routine phytoplankton monitoring.
机译:监测海洋微藻对预测和管理有害藻华非常重要。藻类微阵列检测(MIDTAL)是由FP7资助的欧盟项目,旨在建立多物种微阵列作为辅助监测机构的工具。我们测试了MIDTAL微阵列的不同原型版本是否适合在1年内每月监测Oslofjorden外的采样站的适用性。将来自两个不同版本的MIDTAL芯片的微阵列数据与细胞计数(几种)和定量实时PCR(qPCR;仅假单胞菌属)的结果进行了比较。尽管第2.5代微阵列的结果显示出大量的假阳性信号,但第3.3代微阵列数据通常与显微镜和qPCR数据相关,负责编辑:Philippe Garrigues具有三个重要限制:(1)无法可靠地检测到拟南芥属细胞,可能因为在过滤过程中细胞没有被充分保留,或者在提取过程中细胞没有被溶解,并且由于探针的敏感性低; (2)对于非目标物质浓度高的样品,阵列的灵敏度降低; (3)由于与微阵列上显示的属探针存在1 bp的不匹配,因此未检测到亚历山大假单胞菌发生。尽管存在这些缺点,我们的数据仍显示出整体进展以及MIDTAL阵列的潜力。假单胞菌的案例-区分了两种形态相似的物种,无法通过光学显微镜分离-特别是强调了分子方法(如微阵列)在常规浮游植物监测中的附加值。

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