Department of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine, Kyoto, Japan.
Med Mycol. 2019 Nov 1;57(8):997-1003. doi: 10.1093/mmy/myz003.
The prevalence of azole-resistant Aspergillus fumigatus (ARAF) in Japan is unclear. We aimed to investigate the epidemiology of clinically isolated Aspergillus species and the frequency of azole resistance in Aspergillus species, particularly Aspergillus fumigatus, in the Kyoto and Shiga regions of Japan. Strains of clinically isolated Aspergillus species were prospectively collected from nine acute care hospitals. Species identification was performed by DNA sequence analysis, and all strains were subjected to antifungal susceptibility testing. Sequencing of the Aspergillus cyp51A gene and promoter region and genotyping by short tandem repeats were performed for ARAF isolates. A total of 149 strains were collected, and 130 strains were included for the subsequent analysis after the exclusion of duplicate isolates. The most commonly isolated species was Aspergillus fumigatus, accounting for 43.1% (56 isolates) overall, and seven (12.7%) of 55 strains of A. fumigatus were azole-resistant. Azole-resistance of other Aspergillus species were also found that two (22.2%) of nine strains of A. tubingensis and two (28.6%) of seven strains of A. flavus were azole-resistant. DNA sequence analysis of the ARAF strains revealed that two carried the cyp51A TR34/L98H mutation, one carried G448S, one carried M220I, and three had no relevant mutations (wild type). Genotyping and phylogenetic analyses showed that the TR34/L98H strains were clustered with the strains from the Netherlands and France. These data suggest the emergence of ARAF with TR34/L98H in Japan, and continuous surveillance will be important to identify trends in resistance.
日本的烟曲霉唑耐药（ARAF）流行情况尚不清楚。我们旨在研究日本京都和滋贺地区临床分离的曲霉属物种的流行病学以及曲霉属物种（尤其是烟曲霉）的唑类耐药频率。从 9 家急性护理医院前瞻性采集临床分离的曲霉属菌株。通过 DNA 序列分析进行物种鉴定，所有菌株均进行抗真菌药敏试验。对 ARAF 分离株进行曲霉 CYP51A 基因和启动子区测序和短串联重复序列基因分型。共采集了 149 株菌株，排除重复分离株后，有 130 株菌株纳入后续分析。最常分离的物种是烟曲霉，占总数的 43.1%（56 株），55 株烟曲霉中有 7 株（12.7%）为唑类耐药。还发现了其他曲霉属的唑类耐药，9 株烟管曲霉中有 2 株（22.2%），7 株黄曲霉中有 2 株（28.6%）为唑类耐药。对 ARAF 菌株的 DNA 序列分析显示，2 株携带 CYP51A TR34/L98H 突变，1 株携带 G448S，1 株携带 M220I，3 株无相关突变（野生型）。基因分型和系统发育分析表明，TR34/L98H 株与来自荷兰和法国的菌株聚类。这些数据表明，日本出现了具有 TR34/L98H 的 ARAF，持续监测对于识别耐药趋势非常重要。
