Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria
Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria
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DOI：
10.1002/tox.10108
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发表时间：
2003-04-01
影响因子：
4.5
通讯作者：
Saint, CP
中科院分区：
文献类型：
--
作者：
Fergusson, KM;Saint, CP
对水体进行常规监测。存在潜在毒性的蓝细菌;然而，目前用于确认毒性的方法复杂且昂贵。在这里，我们描述了应用基因为基础的技术，以快速识别产柱孢菌素的蓝藻，特别是，raciborskii spellospermopsis。建立了一种多重聚合酶链反应（PCR）检测方法，可以同时鉴定与柱孢菌素产生相关的聚酮合酶（PKS）和肽合成酶（PS）决定簇，并区分C. raciborskii从其他产圆柱精子蛋白酶的蓝藻物种Anabaenaberatia和Aphanizomenonovalisporum中分离，通过靶向rpoC 1基因。21 C. raciborskii，5A. bernitus，10Aph.用高效液相色谱-质谱联用或基质辅助激光解吸电离飞行时间质谱联用技术对卵孢霉分离物和3个环境样品的PCR结果进行了分析，结果均与其毒理学状态一致，C. raciborskii总是被正确识别。PCR检测是一种快速、可靠和经济的方法，可以评估这些生物体形成的蓝藻水华的毒性潜力。(C)2003 Wiley Periodicals，Inc.
Water bodies are routinely monitored for the. presence of potentially toxic cyanobacteria; however, the methodology for confirming toxicity is currently complex and expensive. Here we describe the application of gene-based technology to rapidly identify cylindrospermopsin-producing cyanobacteria, specifically, Cylindrospermopsis raciborskii. A multiplex polymerase chain reaction (PCR) test was developed that simultaneously identified polyketide synthase (pks) and peptide synthetase (ps) determinants associated with cylindrospermopsin production and distinguished C. raciborskii from other cylindrospermopsin-producing cyanobacteria of the species Anabaena bergii and Aphanizomenon ovalisporum, by targeting the rpoC1 gene. Twenty-one C. raciborskii, 5A. bergii, 10Aph. ovalisporum isolates and 3 environmental samples all yielded PCR results consistent with their toxicological status, as assessed by high-performance liquid chromatography coupled to mass spectrometry or matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, and C. raciborskii was always correctly identified. The PCR test is a rapid, reliable, and economical way of assessing the toxic potential of cyanobacterial blooms formed by these organisms. (C) 2003 Wiley Periodicals, Inc.