Buil Jochem B, Zoll Jan, Verweij Paul E, Melchers Willem J G
Department of Medical Microbiology, Radboud University Medical Center, Nijmegen, Netherlands.
Center of Expertise in Mycology Radboudumc/CWZ, Nijmegen, Netherlands.
Front Microbiol. 2018 Mar 21;9:515. doi: 10.3389/fmicb.2018.00515. eCollection 2018.
diseases are often caused by . Azoles are the mainstay of therapy, but the management of aspergillosis is hampered by the emergence of azole resistance. Rapid detection of azole resistance might benefit treatment outcome by early treatment modifications. However, the yield of fungal culture in invasive aspergillosis is low and susceptibility testing requires several days to be completed. To overcome the low yield of fungal cultures and slow detection of resistance, it is possible to use molecular tools directly on clinical specimens in order to rapidly detect molecular markers of azole resistance. Molecular tools to detect resistant markers in the gene can be expected to be less sensitive compared to molecular tools to detect DNA as the gene is a single copy gene and the target for DNA is often a multi-copy gene. In this mini-review, we summarize the current molecular tools for detection of azole-resistant directly in clinical material. Several in-house PCR assays have been applied directly on clinical material. Furthermore, two assays are commercial available; the AsperGenius and MycoGENIE. The amplification of resistance markers was successful in 70-100% of samples that were positive for DNA in BAL samples using the AsperGenius assay. Despite using several samples per patient, amplification of resistance markers was only successful in 33-57% of patients with DNA in blood. Furthermore, several sequence based methods have been applied with the benefit of the ability to detect other gene alterations.
疾病通常由……引起。唑类药物是治疗的主要手段,但曲霉病的治疗因唑类耐药性的出现而受到阻碍。快速检测唑类耐药性可能通过早期调整治疗方案而改善治疗结果。然而,侵袭性曲霉病中真菌培养的阳性率较低,药敏试验需要数天才能完成。为了克服真菌培养阳性率低和耐药性检测缓慢的问题,可以直接在临床标本上使用分子工具,以便快速检测唑类耐药性的分子标志物。与检测……DNA的分子工具相比,检测……基因中耐药标志物的分子工具可能灵敏度较低,因为……基因是单拷贝基因,而……DNA的靶标通常是多拷贝基因。在本综述中,我们总结了目前直接在临床材料中检测唑类耐药……的分子工具。几种内部PCR检测方法已直接应用于临床材料。此外,有两种检测方法可供商业使用;即AsperGenius和MycoGENIE。使用AsperGenius检测方法,在BAL样本中……DNA呈阳性的样本中,耐药标志物的扩增在70%-100%的样本中成功。尽管每位患者使用了多个样本,但在血液中……DNA呈阳性的患者中,耐药标志物的扩增仅在33%-57%的患者中成功。此外,已经应用了几种基于序列的方法,其优点是能够检测其他……基因改变。