Additional file 1: of A high-throughput sequencing assay to comprehensively detect and characterize unicellular eukaryotes and helminths from biological and environmental samples

Table S1. Examples of genera targeted by each primer pair. Table S2. Primer characteristics of primers from the literature. Table S3. Amplification of positive controls. The table shows the results of the sequencing assay when performed on pools of DNA from known parasites, with and without addition of Anopheles/Escherichia/Human DNA. Each column shows the percentage of the reads that match each observed species. Green text represents on-target species and red text shows off-target (often host) species amplification. Table S4. Results from all samples. Explanation of each column is presented in the excel sheet. Figure S1. Complementarity of the primer pairs targeting the same taxonomic groups. Apicomplexa and Nematoda each required three primer pairs to capture the diversity within these groups. The taxa amplified by each of the three primer sets are presented as Venn diagrams showing the overlap in species coverage. The percent within each sector is shown in parentheses. Figure S2. Overview of the pipeline for the in silico assessment of the primer amplification range, information content and specificity. Figure S3. PrimerTree results for each newly designed primer. The figures show, for each primer set, the PrimerTree plot and amplicon lengths. The PrimerTree results were restricted only to the targeted group to show the diversity of on-target taxonomic groups amplifiable. (ZIP 2497 kb)

表S1：各引物对靶向的属类示例。
表S2：文献来源引物的引物特征参数。
表S3：阳性对照扩增结果。该表格展示了针对已知寄生虫DNA混合池开展测序检测的结果，设置了添加与不添加按蚊/埃希氏菌/人类DNA的两组对照。每一列对应匹配各观测物种的测序读段占比。绿色字体代表靶标物种，红色字体代表脱靶（通常为宿主）物种的扩增结果。
表S4：全部样本检测结果。各列的详细说明请见附带的Excel表格。
图S1：靶向同一分类群的引物对互补性分析。顶复门（Apicomplexa）与线虫动物门（Nematoda）各需3组引物对以覆盖该类群内的多样性。3组引物各自扩增的分类群以韦恩图展示物种覆盖范围的重叠情况，扇区内的百分比以括号标注。
图S2：引物扩增范围、信息含量与特异性的计算机模拟评估流程概览。
图S3：每组新设计引物的PrimerTree分析结果。该图针对每组引物集展示了PrimerTree绘图与扩增子长度。本次PrimerTree分析仅限定于靶标类群，以展示可扩增的靶标分类群多样性。（压缩包大小：2497 KB）