Department of Internal Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea.
Department of Internal Medicine, Yonsei University College of Medicine, Seoul, Korea.
Yonsei Med J. 2020 Aug;61(8):698-704. doi: 10.3349/ymj.2020.61.8.698.
With changing fungal epidemiology and azole resistance in species, identifying fungal species and susceptibility patterns is crucial to the management of aspergillosis and mucormycosis. The objectives of this study were to evaluate performance of panfungal polymerase chain reaction (PCR) assays on formalin-fixed paraffin embedded (FFPE) samples in the identification of fungal species and in the detection of azole-resistance mutations in the gene at a South Korean hospital.
A total of 75 FFPE specimens with a histopathological diagnosis of aspergillosis or mucormycosis were identified during the 10-year study period (2006-2015). After deparaffinization and DNA extraction, panfungal PCR assays were conducted on FFPE samples for fungal species identification. The identified fungal species were compared with histopathological diagnosis. On samples identified as , sequencing to identify frequent mutations in the gene [tandem repeat 46 (TR46), L98H, and M220 alterations] that confer azole resistance was performed.
Specific fungal DNA was identified in 31 (41.3%) FFPE samples, and of these, 16 samples of specific fungal DNA were in accord with a histopathological diagnosis of aspergillosis or mucormycosis; 15 samples had discordant histopathology and PCR results. No azole-mediating gene mutation was noted among nine cases of aspergillosis. Moreover, no mutations were identified among three cases with history of prior azole use.
Panfungal PCR assay with FFPE samples may provide additional information of use to fungal species identification. No azole-resistance mediating mutations in the gene were identified among FFPE samples during study period.
随着 属真菌的流行病学和唑类耐药性的变化,确定真菌种类和药敏模式对于曲霉病和毛霉病的治疗至关重要。本研究旨在评估在韩国一家医院中,使用基于聚合酶链反应(PCR)的泛真菌检测方法对福尔马林固定石蜡包埋(FFPE)样本进行真菌种类鉴定和 基因中唑类耐药突变检测的性能。
在 10 年的研究期间(2006-2015 年),共从 75 例组织病理学诊断为曲霉病或毛霉病的 FFPE 标本中鉴定出真菌。脱蜡和 DNA 提取后,对 FFPE 样本进行泛真菌 PCR 检测以鉴定真菌种类。将鉴定出的真菌种类与组织病理学诊断进行比较。在鉴定为 的样本中,对 基因(串联重复 46 [TR46]、L98H 和 M220 改变)中常见耐药突变进行测序。
在 31 例(41.3%)FFPE 样本中鉴定出特定真菌 DNA,其中 16 例特定真菌 DNA 与曲霉病或毛霉病的组织病理学诊断一致;15 例样本的组织病理学和 PCR 结果不一致。在 9 例曲霉病中未发现唑类药物介导的 基因突变。此外,在有既往唑类药物使用史的 3 例病例中未发现 突变。
使用 FFPE 样本进行泛真菌 PCR 检测可能提供有助于真菌种类鉴定的额外信息。在研究期间,未从 FFPE 样本中鉴定出 基因中的唑类耐药介导突变。