Quality Controls.

aOne region of an 8 gasket PTP was used for a positive control (C). Negatives, positives, and pilot samples (representing a subset of three subjects) were sequenced in one region on the same run, and pilot l (A) and negative control (B) sequences partitioned by MID.bQIIME restrictions: Minimum length = 100 (after trimming forward primer and MID); maximum “N” = 1, maximum homocopolymer = 10; maximum forward primer mismatch = 2; maximum barcode mismatch = 2.cGenus assignments (sequence counts): Unclassified fungi (6 sequences); Saccharomyces (3); Tumularia (3); Malassezia (2); Rhodotorula (2); Candida (1); Ceratobasidium (1); Galerina (1). One of the genera assignments (Ceratobasidium) was at a very weak E-value (3.9); all others were at very strong E-values (−85 to −177).dIncludes Candida. All non-Candida genera are constituted by singleton sequences; 4 (Scutellospora,Tomentella, Saccharomyces, Cryptococcus) have very weak E-values (0.11–4.8); 1 (Fusarium) has a strong E-value (−119).a. 本实验采用8垫片PTP的一个区域作为阳性对照（C）。阴性对照、阳性对照及代表3名受试对象子集的预实验样本，于同一次测序运行中在同一区域完成测序；其中预实验样本l（A）与阴性对照（B）的序列通过MID（分子标识符，Molecular Identifier）进行划分。 b. QIIME（定量微生物生态学分析软件，Quantitative Insights Into Microbial Ecology）质控过滤阈值设置如下：修剪正向引物与MID后的序列最小长度为100；最大允许模糊碱基（N）数为1；最大允许同聚物长度为10；正向引物最大错配数为2；测序条形码最大错配数为2。 c. 属水平分类注释及对应序列计数如下：未分类真菌（6条序列）、酿酒酵母属（Saccharomyces，3条）、Tumularia（3条）、马拉色菌属（Malassezia，2条）、红酵母属（Rhodotorula，2条）、假丝酵母属（Candida，1条）、角担菌属（Ceratobasidium，1条）、盔孢伞属（Galerina，1条）。其中角担菌属的分类注释对应的E-值（期望分值，Expectation Value）为3.9（极低），其余所有分类注释的E-值均处于-85至-177的强负值区间。 d. 本次分析纳入假丝酵母属（Candida）。所有非假丝酵母属的分类单元均由单例序列构成；其中4个属（球囊霉属Scutellospora、Tomentella、酿酒酵母属Saccharomyces、隐球菌属Cryptococcus）对应的E-值极弱（0.11~4.8），1个属（镰孢菌属Fusarium）对应的E-值较强（-119）。