Jan Hao-En, Tsai Chin-Shiang, Cia Cong-Tat, Lee Ching-Chi, Chen Ying-Wen, Lee Nan-Yao, Li Chia-Wen, Li Ming-Chi, Syue Ling-Shan, Lo Ching-Lung, Chang Tsung-Chain, Wu Chi-Jung, Ko Wen-Chien, Chen Po-Lin
Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan 701, Taiwan.
Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan 701, Taiwan.
J Fungi (Basel). 2024 Jul 29;10(8):530. doi: 10.3390/jof10080530.
A fungal polymerase chain reaction (PCR) amplifies conserved genes across diverse species, combined with the subsequent hybridization of amplicons using a specific oligonucleotide microarray, allowing for the rapid detection of pathogens at the species level. However, the performance of microarrays in diagnosing invasive mold infections (IMI) from infected tissue samples is rarely reported. During the 4-year study period, all biopsied tissue samples from patients with a suspected IMI sent for microarray assays were analyzed. A partial segment of the internal transcribed spacer (ITS) region was amplified by nested PCR after DNA extraction. Amplicons were hybridized with specific probes for a variety of mold species using an in-house oligonucleotide microarray. A total of 80 clinical samples from 74 patients were tested. A diagnosis of an IMI was made in 10 patients (4 proven, 1 probable, 3 possible, 2 clinical suspicion). The PCR/microarray test was positive for three out of four proven IMIs, one probable IMI, and one out of three possible IMIs. Two patients with positive PCR/microarray findings were considered to have clinical suspicion of an IMI, and their responsible physicians initiated antifungal therapy despite the absence of supporting microbiological and histological evidence. Clinical diagnoses were categorized into non-IMI and IMI groups (including proven, probable, possible, and clinical suspicion). The sensitivity and specificity of the microarray in diagnosing the IMIs were 70% and 95.7%, respectively, while the sensitivity and specificity of the culture and histological findings were 10%/96.3% and 40.0%/100%, respectively. PCR-based methods provide supportive microbiological evidence when culture results are inconclusive. The combination of a microarray with fungal culture and histology promotes the precise diagnosis of IMIs in difficult-to-diagnose patients.
真菌聚合酶链反应（PCR）可扩增不同物种间的保守基因，并结合随后使用特定寡核苷酸微阵列对扩增子进行杂交，从而能够在物种水平上快速检测病原体。然而，关于微阵列在从感染组织样本中诊断侵袭性霉菌感染（IMI）方面的性能报道很少。在为期4年的研究期间，对所有因疑似IMI而送检进行微阵列检测的患者的活检组织样本进行了分析。DNA提取后，通过巢式PCR扩增内部转录间隔区（ITS）区域的部分片段。使用自制的寡核苷酸微阵列将扩增子与针对多种霉菌物种的特异性探针进行杂交。共检测了来自74例患者的80份临床样本。10例患者被诊断为IMI（4例确诊，1例很可能，3例可能，2例临床疑似）。PCR/微阵列检测在4例确诊的IMI中有3例呈阳性，1例很可能的IMI呈阳性，3例可能的IMI中有1例呈阳性。2例PCR/微阵列结果呈阳性的患者被认为临床疑似IMI，尽管缺乏支持性的微生物学和组织学证据，但其责任医师仍启动了抗真菌治疗。临床诊断分为非IMI组和IMI组（包括确诊、很可能、可能和临床疑似）。微阵列诊断IMI的敏感性和特异性分别为70%和95.7%，而培养和组织学检查结果的敏感性和特异性分别为10%/96.3%和40.0%/100%。当培养结果不明确时，基于PCR的方法可提供支持性的微生物学证据。微阵列与真菌培养及组织学检查相结合，有助于对难以诊断的患者进行IMI的精确诊断。
