Laboratory of Environmental Microbiology and Toxicology, School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, Hong Kong, People's Republic of China.
Institute for Advanced Study, Shenzhen University, Shenzhen, 518060, People's Republic of China.
Appl Microbiol Biotechnol. 2018 Jul;102(14):5859-5871. doi: 10.1007/s00253-018-9078-2. Epub 2018 May 25.
Research on anammox (anaerobic ammonium oxidizing) bacteria is important due to their biogeochemical and industrial application significance since the first discovery made over two decades ago. By coupling NH and NO biochemically to form N gas, anammox bacteria contribute significantly to global marine and terrestrial nitrogen balance (responsible for 50, 940, and 437% of the nitrogen loss for marine, lakes, and paddy soil) and are also useful in energy-conserving nitrogen removal in wastewater treatment. PCR-based detection and quantification of anammox bacteria are an easy, essential, and widely accessible technique used ubiquitously for studying them in many environmental niches. In this article, we make a summary on practical applications of 16S rRNA and functional gene PCR primers, including hydrazine dehydrogenase (Hzo), nitrite reductase (NirS), hydrazine synthase (Hzs), and cytochrome c biogenesis proteins (Ccs) in detection of them. PCR primer performances in both practical applications and tests in silico are also presented for comparison. For detecting general and specific anammox bacterial groups, selection of appropriate PCR primers for different environmental samples and practical application guidance on choice of appropriate primer pairs for different purposes are also offered. This article provides practical information on selection and application of PCR technique in detection of anammox bacteria from the diverse environments to further promote convenient applications of this technique in research and other purposes.
由于自二十多年前首次发现以来，厌氧氨氧化（anaerobic ammonium oxidizing）细菌在生物地球化学和工业应用方面具有重要意义，因此对其开展研究非常重要。通过将 NH 和 NO 生物化学偶联形成 N 气体，厌氧氨氧化细菌对全球海洋和陆地氮平衡做出了重大贡献（分别占海洋、湖泊和稻田氮损失的 50%、940%和 437%），并且在废水处理中的节能脱氮方面也非常有用。基于 PCR 的厌氧氨氧化细菌检测和定量是一种简单、基本且广泛应用的技术，可用于研究许多环境小生境中的厌氧氨氧化细菌。在本文中，我们总结了 16S rRNA 和功能基因 PCR 引物的实际应用，包括肼脱氢酶（Hzo）、亚硝酸盐还原酶（NirS）、肼合酶（Hzs）和细胞色素 c 生物发生蛋白（Ccs）在检测中的应用。还介绍了 PCR 引物在实际应用和计算机模拟测试中的性能，以便进行比较。为了检测一般和特定的厌氧氨氧化细菌群体，我们还为不同环境样本选择了合适的 PCR 引物，并提供了针对不同目的选择合适引物对的实用指导。本文提供了从不同环境中选择和应用 PCR 技术检测厌氧氨氧化细菌的实用信息，以进一步促进该技术在研究和其他目的中的便捷应用。
