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使用基于定量聚合酶链反应(qPCR)的检测方法监测休闲水域中有毒的卵形卡盾藻(Ostreopsis cf. ovata)。

Monitoring toxic Ostreopsis cf. ovata in recreational waters using a qPCR based assay.

作者信息

Casabianca Silvia, Perini Federico, Casabianca Anna, Battocchi Cecilia, Giussani Valentina, Chiantore Mariachiara, Penna Antonella

机构信息

Section of Environmental Biology, Dep. of Biomolecular Sciences, University of Urbino, Viale Trieste 296, 61121 Pesaro, Italy.

Department of Earth, Environment and Life Sciences, University of Genoa, Corso Europa 26, 16132 Genoa, Italy.

出版信息

Mar Pollut Bull. 2014 Nov 15;88(1-2):102-9. doi: 10.1016/j.marpolbul.2014.09.018. Epub 2014 Oct 1.

Abstract

Ostreopsis sp. is a toxic marine benthic dinoflagellate that causes high biomass blooms, posing a threat to human health, marine biota and aquaculture activities, and negatively impacting coastal seawater quality. Species-specific identification and enumeration is fundamental because it can allow the implementation of all the necessary preventive measures to properly manage Ostreopsis spp. bloom events in recreational waters and aquaculture farms. The aim of this study was to apply a rapid and sensitive qPCR method to quantify Ostreopsis cf. ovata abundance in environmental samples collected from Mediterranean coastal sites and to develop site-specific environmental standard curves. Similar PCR efficiencies of plasmid and environmental standard curves allowed us to estimate the LSU rDNA copy number per cell. Moreover, we assessed the effectiveness of mitochondrial COI and cob genes as alternative molecular markers to ribosomal genes in qPCR assays for Ostreopsis spp. quantification.

摘要

链状裸甲藻是一种有毒的海洋底栖甲藻,会引发高生物量水华,对人类健康、海洋生物群落和水产养殖活动构成威胁,并对沿海水质产生负面影响。物种特异性鉴定和计数至关重要,因为这样可以实施所有必要的预防措施,以妥善管理娱乐水域和水产养殖场中的链状裸甲藻水华事件。本研究的目的是应用一种快速灵敏的定量聚合酶链反应(qPCR)方法,对从地中海沿岸站点采集的环境样本中的卵形链状裸甲藻丰度进行定量,并绘制特定站点的环境标准曲线。质粒和环境标准曲线相似的聚合酶链反应效率使我们能够估计每个细胞的大亚基核糖体DNA(LSU rDNA)拷贝数。此外,我们评估了线粒体细胞色素氧化酶亚基I(COI)基因和细胞色素b(cob)基因作为定量聚合酶链反应分析中核糖体基因的替代分子标记,用于链状裸甲藻物种定量的有效性。

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