Overview of PCR Methods Applied for the Identification of Freshwater Toxigenic Cyanobacteria

Jian Yuan, K. Yoon
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引用次数: 3

Abstract

Although cyanobacteria are essential microorganisms on earth, some cyanobacteria produce toxins known as cyanotoxins, threatening humans and animals’ health. Hence, it is imperative to rapidly and accurately identify those toxic cyanobacteria. Unfortunately, traditional microscopic methods have limitations for accurate identification due to the lack of discernable morphological difference between toxic and non-toxic strains within the same cyanobacterial species or genus. In contrast, their genetic profiles are inherently conserved; therefore, nucleic acid-based assays can be more reliable for precise identification. Furthermore, molecular assays can provide high throughput and significantly reduce the turnaround time of test results. Such advantages make those assays a preferred method for rapid detection and early warning of potential toxicity. Toxigenic cyanobacterial species have synthetase genes (DNAs) for toxin production, which can be excellent marker genes. Numerous molecular assays targeting cyanotoxin synthetase genes have been developed for the identification of toxigenic cyanobacteria at various taxonomic levels. Polymerase chain reaction (PCR)-based assays are the most prevailing. Among different versions of PCR assays, the real-time quantitative PCR can be utilized to quantify the genes of interest in samples, fulfilling the purpose of both taxonomic recognition and biomass estimation. Reverse transcription (RT)-PCR assays can be used to detect transcripts (i.e., mRNAs) from toxin synthetase genes, probably enhancing the predictive value of PCR detection for toxin production from observed cyanobacterial species. Nevertheless, the utility of toxin synthetase gene- or its transcript-based PCR assays for routine cyanotoxin monitoring needs to be further evaluated on a large scale.
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淡水产毒蓝藻PCR鉴定方法综述
虽然蓝藻是地球上必不可少的微生物,但一些蓝藻会产生被称为蓝藻毒素的毒素,威胁着人类和动物的健康。因此,必须快速准确地识别这些有毒的蓝藻。不幸的是,由于在同一蓝藻物种或属内缺乏有毒和无毒菌株之间可识别的形态学差异,传统的显微方法具有准确鉴定的局限性。相比之下,他们的基因图谱是固有的保守;因此,基于核酸的检测可以更可靠地进行精确鉴定。此外,分子分析可以提供高通量,并显著减少测试结果的周转时间。这些优点使这些测定法成为快速检测和早期预警潜在毒性的首选方法。产毒蓝藻具有产毒合成酶基因(dna),是很好的标记基因。许多针对蓝藻毒素合成酶基因的分子分析已经开发出来,用于在不同的分类水平上鉴定产毒素蓝藻。聚合酶链反应(PCR)为基础的分析是最普遍的。在不同版本的PCR检测中,实时定量PCR可以用来定量样品中感兴趣的基因,实现分类识别和生物量估算的目的。逆转录(RT)-PCR检测可用于检测毒素合成酶基因的转录本(即mrna),这可能提高了PCR检测对观察到的蓝藻物种毒素产生的预测价值。然而,毒素合成酶基因或其转录为基础的PCR检测在常规蓝藻毒素监测中的应用需要进一步大规模评估。
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