High-throughput sequencing of 16S rRNA amplicons is an effective method for diversity analysis of anammox bacteria. However, the corresponding taxonomic database and suitable cut-off value are unavailable for processing high-throughput sequencing data. This study is the first to construct a taxonomic database, comprising 17 sequences of enriched anammox strains (regarded as seed sequences) and 2925 uncultured sequences from environmental samples. Moreover, a suitable cut-off value was determined based on pairwise evolutionary distances between the 16S rRNA gene sequences of the 17 identified species and diversity indexes of community in the taxonomic database. Furthermore, the proposed method was validated with multiple environmental samples. In summary, the taxonomic database and proposed cut-off value targeting the 16S rRNA gene presented in this study can be effectively employed to explore the diversity of anammox bacteria.

16S rRNA扩增子的高通量测序是厌氧氨氧化细菌多样性分析的有效方法。然而，没有相应的分类数据库和合适的截止值来处理高通量测序数据。本研究首次构建了分类数据库，包含17个富集厌氧氨氧化菌株序列（视为种子序列）和来自环境样本的2925个未培养序列。此外，根据17个已鉴定物种的16S rRNA基因序列之间的成对进化距离和分类数据库中群落的多样性指数，确定了合适的截止值。此外，所提出的方法通过多个环境样本进行了验证。总之，本研究中提出的针对 16S rRNA 基因的分类数据库和建议的临界值可以有效地用于探索厌氧氨氧化细菌的多样性。