Real-time qPCR screening was performed for comparative analysis of fungal distribution between AD patients and healthy individuals. To perform amplification, we used pan-fungal primers (Table 1)10 (link)11 (link)12 (link), which are universally selected for targeting the fungal ITS1 region. Real-time qPCR was conducted with iTaq Universal SYBR Green PCR master mix (Bio-Rad, Hercules, CA, USA) according to the manufacturer’s instructions on a CFX96 real-time PCR detection system. The amplicons were purified using a PCR purification kit (Qiagen, Valencia, CA, USA) according to the manufacturer’s instructions. The purified amplicons were sequenced using BigDye® Terminator V3.1 cycle sequencing kit (Applied Biosystems, Foster City, CA, USA) with forward and reverse primers initially used in PCR amplification. To ascertain species identity, the characterized sequence was analyzed using BLASTn to align sequence similarity with intended target genes in NCBI (http://blast.ncbi.nlm.nih.gov/Blast.cgi).