Departamento de Ciências Biológicas, ESALQ, Universidade de São Paulo, Av. Pádua Dias 11, 13418-900, Piracicaba, São Paulo, Brasil.
Environ Toxicol. 2012 Aug;27(8):461-71. doi: 10.1002/tox.20659. Epub 2010 Nov 29.
The aim of this study was to evaluate the use of new oligonucleotide primers (mcyB-F/R, mcyB-F/R-A, and mcyB-F/R-B) designed from Brazilian cyanobacteria for the detection of microcystin-producing genotypes in 27 environmental samples from water reservoirs and 11 strains of Microcystis. Microcystins were found using HPLC in all 11 strains and 19 of the environmental samples. The new oligonucleotide primers amplified fragments of microcystin-producing genes, including the eight environmental samples in which no microcystins were detected by HPLC, but which presented amplified fragments, thereby demonstrating the existence of microcystin-producing genes. The new oligonucleotide primers exhibited better specificity when used with environmental samples and were more reliable in comparison with those described in the literature (mcyB-FAA/RAA and mcyA-Cd/FR), which generate false-negative results. The better performance of these new oligonucleotide primers underline the need for designing molecular markers that are well fitted to the regional biological diversity. As this is a fast predictive technique for determining the presence or absence of microcystins, it could be used either alone or in conjunction with other techniques, such as the screening of samples to be sent for quantitative toxicological analysis using HPLC, thereby reducing monitoring cost and time. © 2010 Wiley Periodicals, Inc. Environ Toxicol, 2012.
本研究的目的是评估从巴西蓝藻设计的新寡核苷酸引物(mcyB-F/R、mcyB-F/R-A 和 mcyB-F/R-B)在检测 27 个水库环境样本和 11 株微囊藻中的产微囊藻毒素基因型中的应用。所有 11 株菌株和 19 个环境样本均使用 HPLC 检测到微囊藻毒素。新的寡核苷酸引物扩增了产微囊藻毒素基因的片段,包括 8 个环境样本,这些样本用 HPLC 检测不到微囊藻毒素,但却有扩增片段,从而证明存在产微囊藻毒素基因。与文献中描述的寡核苷酸引物(mcyB-FAA/RAA 和 mcyA-Cd/FR)相比,新的寡核苷酸引物在检测环境样本时表现出更好的特异性,并且更可靠,因为后者会产生假阴性结果。这些新的寡核苷酸引物具有更好的性能,这突显了设计适合于区域生物多样性的分子标记的必要性。由于这是一种快速预测技术,用于确定微囊藻毒素的存在与否,因此可以单独使用或与其他技术结合使用,例如对要进行 HPLC 定量毒理学分析的样本进行筛选,从而降低监测成本和时间。 © 2010 Wiley Periodicals, Inc. Environ Toxicol, 2012.