Resendiz-Sharpe Agustin, Van Holm Wannes, Merckx Rita, Pauwels Martine, Teughels Wim, Lagrou Katrien, Vande Velde Greetje
Department of Imaging and Pathology, Biomedical MRI, KU Leuven, 3000 Leuven, Belgium.
Department of Oral and Health Sciences, KU Leuven, 3000 Leuven, Belgium.
J Fungi (Basel). 2022 Oct 25;8(11):1120. doi: 10.3390/jof8111120.
Increasing resistance to triazole antifungals in is worrisome because of the associated high mortality of triazole-resistant (TRAF) infections. While most studies have focused on single triazole-susceptible (WT) or TRAF infections, reports of TRAF cases developing mixed WT and TRAF infections have been described in several studies. However, the prevalence of mixed infections and their responses to current recommended therapies are unknown and could be inappropriate, leading to poor clinical outcomes. To address the urgent need for tools to diagnose, monitor disease development and therapy efficacies in mixed infection settings where quantification of WT versus TRAF is key, this study developed a novel qPCR assay to differentiate WT and TRAF harboring the -TR/L98H mutation. The proposed assay successfully quantified and discriminated TRAF-TR in vitro and in vivo, which was achieved by increasing the yield of extracted DNA through improved homogenization and specific primers targeting the WT-sequence or TR-insertion and a TaqMan-probe directed to . The here-developed qPCR assay overcomes sensitivity issues of methodologies such as CFU counts, providing specific, reproducible, and reliable quantitative information to study and follow up the (interplay and individual) effects of mixed infections on disease development and treatment responses.
曲霉菌对三唑类抗真菌药物的耐药性不断增加令人担忧，因为耐三唑类曲霉菌（TRAF）感染的相关死亡率很高。虽然大多数研究集中在单一的三唑类敏感（WT）或TRAF感染上，但在几项研究中已经描述了TRAF病例发展为WT和TRAF混合感染的报告。然而，混合感染的患病率及其对当前推荐治疗的反应尚不清楚，可能并不合适，从而导致临床结果不佳。为了满足在WT与TRAF定量至关重要的混合感染环境中诊断、监测疾病发展和治疗效果的工具的迫切需求，本研究开发了一种新型qPCR检测方法，以区分携带-TR/L98H突变的WT和TRAF。所提出的检测方法在体外和体内成功地对TRAF-TR进行了定量和区分，这是通过改进匀浆提高提取DNA的产量、靶向WT序列或TR插入的特异性引物以及针对的TaqMan探针实现的。这里开发的qPCR检测方法克服了诸如CFU计数等方法的灵敏度问题，为研究和跟踪混合曲霉菌感染对疾病发展和治疗反应（相互作用和个体）的影响提供了具体、可重复和可靠的定量信息。
