Koc University Is Bank Center for Infectious Disease (KUISCID), Istanbul, Turkey.
Koç University, Graduate School of Health Science, Medical Microbiology Department, Istanbul, Turkey.
Microb Drug Resist. 2024 May;30(5):210-213. doi: 10.1089/mdr.2023.0198. Epub 2024 Feb 12.
There is an emerging fluconazole resistance in in recent years. The leading mechanism causing azole resistance in is the Y132F codon alteration in the ERG11 gene which encodes the target enzyme of azole drugs. In this study, we evaluated the sensitivity, compatibility, and specificity of a novel tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) method for rapid detection of the Y132F mutation in fluconazole nonsusceptible Antifungal susceptibility tests for detection of fluconazole resistance were performed by broth microdilution according to the CLSI guidelines. All susceptible and nonsusceptible isolates were analyzed for ERG11 mutations with Sanger sequencing. T-ARMS-PCR was fully concordant with the Sanger sequencing (100% of sensitivity and specificity) for detection of Y132F mutations. T-ARMS-PCR method could be a rapid, simple, accurate, and economical assay in the early detection of the most common cause of fluconazole resistance in isolates. In routine laboratories with high isolation rates, performing the T-ARMS-PCR for early detection of the most common reason of fluconazole resistance in sis, could be a life-saving approach for directing antifungal therapy before obtaining the definitive antifungal susceptibility tests results.
近年来, 中出现了一种新的氟康唑耐药性。导致唑类药物耐药的主要机制是 ERG11 基因中的 Y132F 密码子改变,该基因编码唑类药物的靶酶。在本研究中,我们评估了一种新的四引物扩增受阻突变系统-聚合酶链反应(T-ARMS-PCR)方法检测氟康唑不敏感 中 Y132F 突变的敏感性、兼容性和特异性。根据 CLSI 指南,通过肉汤微量稀释法进行抗真菌药敏试验检测氟康唑耐药性。对所有敏感和不敏感的 分离株进行 Sanger 测序分析 ERG11 突变。T-ARMS-PCR 与 Sanger 测序完全一致(敏感性和特异性均为 100%),用于检测 Y132F 突变。T-ARMS-PCR 方法可用于早期快速、简单、准确、经济地检测 中最常见的氟康唑耐药原因。在高 分离率的常规实验室中,进行 T-ARMS-PCR 以早期检测 中最常见的氟康唑耐药原因,在获得明确的抗真菌药敏试验结果之前,可能是指导抗真菌治疗的一种救命方法。