CYANOCHIP: An Antibody Microarray for High-Taxonomical-Resolution Cyanobacterial Monitoring

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• 作者：Yolanda Blanco ; Antonio Quesada ; Ignacio Gallardo-Carre帽o ; Jacobo Aguirre ; Victor Parro
• 刊名：Environmental Science & Technology
• 出版年：2015
• 出版时间：February 3, 2015
• 年：2015
• 卷：49
• 期：3
• 页码：1611-1620
• 全文大小：483K
• ISSN：1520-5851

文摘

Cyanobacteria are Gram-negative photosynthetic prokaryotes that are widespread on Earth. Eutrophication and global warming make some aquatic ecosystems behave as bioreactors that trigger rapid and massive cyanobacterial growth with remarkable economic and health consequences. Rapid and efficient early warning systems are required to support decisions by water body authorities. We have produced 17 specific antibodies to the most frequent cyanobacterial strains blooming in freshwater ecosystems, some of which are toxin producers. A sandwich-type antibody microarray immunoassay (CYANOCHIP) was developed for the simultaneous testing of any of the 17 strains, or other closely related strains, in field samples from different habitats (water, rocks, and sediments). We titrated and tested all of the antibodies in succession using a fluorescent sandwich microarray immunoassay. Although most showed high specificity, we applied a deconvolution method based on graph theory to disentangle the few existing cross-reactions. The CYANOCHIP sensitivity ranged from 10² to 10⁴ cells mL^鈥?, with most antibodies detecting approximately 10² cells mL^鈥?. We validated the system by testing multiple isolates and crude natural samples from freshwater reservoirs and rocks, both in the laboratory and by in situ testing in the field. The results demonstrated that CYANOCHIP is a valuable tool for the sensitive and reliable detection of cyanobacteria for early warning and research purposes.