Dalyan Cilo Burcu, Topaç Tuncay, Ağca Harun, Sağlam Sezcan, Efe Kadir, Ener Beyza
Uludağ University Faculty of Medicine, Department of Medical Microbiology, Bursa, Turkey.
Mikrobiyol Bul. 2018 Jan;52(1):35-48. doi: 10.5578/mb.63991.
Candida species are among the top 10 pathogens causing bloodstream infections associated with high morbidity, mortality. In spite of the development of new antifungal drugs, epidemiological studies have shown that resistance to antifungal drugs among Candida isolates is becoming a serious problem. The aim of this study was to compare the antifungal broth microdilution methods of the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) for amphotericin B, fluconazole, itraconazole, voriconazole, posaconazole and anidulafungin susceptibility of Candida blood isolates. The study consisted of 74 Candida albicans, 67 Candida parapsilosis, 30 Candida glabrata, and 18 Candida tropicalis isolates. The minimum inhibitory concentrations were determined after 24 and 48 hour of incubation with CLSI method and only 24 hour of incubation with EUCAST method except anidulofungin. The MIC values obtained by both methods were considered to be compatible within ± 2 dilution limits. The categorical agreement between methods for each antifungal agent was assessed using clinical break points and epidemiological cut-off values. The agreement (± 2 dilution) between the methods was found to be species, drug, and incubation time dependent. After 24 hour incubation, good agreement category (> 90%) was detected between amphotericin B, itraconazole, posaconazole and anidulofungin, but was lower category (< 85%) was determined with fluconazole and voriconazole especially for relatively slow growing C.glabrata and C.parapsilosis isolates. Excellent categorical agreement (100%) was observed for amfoterisin B/C.parapsilosis, C.glabrata, C.tropicalis and anidulofungin/C.albicans, C.glabrata, C.tropicalis but least category was determined for posaconazole and C.albicans (71.6% at 24 hour; 73% at 48 hour). In vitro resistance of therapeutically used fluconazole and anidulafungin determined by both methods was rare among C.albicans (1.3%, 2.7% respectively), C.glabrata (0%, 3.3% respectively) and C.tropicalis (0%, 5.6% respectively) isolates but, an increase of non-susceptible isolates were observed among C.parapsilosis (11.9% at 24 hour of incubation; 17.9% at 48 hour of incubation) for fluconazole. There was also a cross resistance between fluconazole and voriconazole for three C.parapsilosis isolates and one multidrug resistant (fluconazole, itraconazole, posaconazole and anidulofungin) C.albicans isolate (fluconazole, itraconazole, posaconazole and anidulofungin). As a result in this study, it was determined thatboth methods were similar and can be used according to preference of laboratories. The CLSI antifungal susceptibility test results can be assessed at the end of 24 hour incubation, but sometimes it is important that the evaluation should be performed as a result of 48 hour incubation in slow growing species such as C.glabrata.
念珠菌属是导致血流感染的十大病原体之一,与高发病率和死亡率相关。尽管开发了新的抗真菌药物,但流行病学研究表明,念珠菌分离株对抗真菌药物的耐药性正成为一个严重问题。本研究的目的是比较临床和实验室标准协会(CLSI)和欧洲抗菌药物敏感性试验委员会(EUCAST)针对两性霉素B、氟康唑、伊曲康唑、伏立康唑、泊沙康唑和阿尼芬净对念珠菌血培养分离株的抗真菌肉汤微量稀释法。该研究包括74株白色念珠菌、67株近平滑念珠菌、30株光滑念珠菌和18株热带念珠菌分离株。采用CLSI方法在孵育24小时和48小时后测定最低抑菌浓度,采用EUCAST方法(阿尼芬净除外)仅在孵育24小时后测定。两种方法获得的最低抑菌浓度值在±2倍稀释限度内被认为是一致的。使用临床断点和流行病学截断值评估每种抗真菌药物方法之间的分类一致性。发现方法之间的一致性(±2倍稀释)取决于菌种、药物和孵育时间。孵育24小时后,两性霉素B、伊曲康唑、泊沙康唑和阿尼芬净之间检测到良好的一致性类别(>90%),但氟康唑和伏立康唑的一致性类别较低(<85%),特别是对于生长相对缓慢的光滑念珠菌和近平滑念珠菌分离株。两性霉素B/近平滑念珠菌、光滑念珠菌、热带念珠菌以及阿尼芬净/白色念珠菌、光滑念珠菌、热带念珠菌观察到极好的分类一致性(100%),但泊沙康唑和白色念珠菌的一致性类别最低(24小时时为71.6%;48小时时为73%)。两种方法测定的治疗用氟康唑和阿尼芬净的体外耐药性在白色念珠菌(分别为1.3%、2.7%)、光滑念珠菌(分别为0%、3.3%)和热带念珠菌(分别为0%、5.6%)分离株中很少见,但在近平滑念珠菌中观察到不敏感分离株有所增加(孵育24小时时为11.9%;孵育48小时时为17.9%)。三株近平滑念珠菌分离株和一株多重耐药(氟康唑、伊曲康唑、泊沙康唑和阿尼芬净)白色念珠菌分离株(氟康唑、伊曲康唑、泊沙康唑和阿尼芬净)中还存在氟康唑和伏立康唑之间的交叉耐药。本研究结果表明,两种方法相似,可根据实验室偏好使用。CLSI抗真菌药敏试验结果可在孵育24小时结束时评估,但对于生长缓慢的菌种如光滑念珠菌来说,有时在孵育48小时后进行评估很重要。