Mohammad Asadzadeh, Suhail Ahmad, Wadha Alfouzan, Inaam Al-Obaid, Bram Spruijtenburg, Eelco F. J. Meijer, Jacques F. Meis, Eiman Mokaddas
{"title":"评估 Etest 和 MICRONAUT-AM 检测法对白色念珠菌的抗真菌药敏试验:MICRONAUT-AM 低估了氟康唑的耐药性,Etest 高估了两性霉素 B 的耐药性","authors":"Mohammad Asadzadeh, Suhail Ahmad, Wadha Alfouzan, Inaam Al-Obaid, Bram Spruijtenburg, Eelco F. J. Meijer, Jacques F. Meis, Eiman Mokaddas","doi":"10.3390/antibiotics13090840","DOIUrl":null,"url":null,"abstract":"Multidrug-resistant Candida auris has recently caused major outbreaks in healthcare facilities. Rapid and accurate antifungal susceptibility testing (AST) of C. auris is crucial for proper management of invasive infections. The Commercial Sensititre Yeast One and Vitek 2 methods underestimate or overestimate the resistance of C. auris to fluconazole and amphotericin B (AMB). This study evaluated the AST results of C. auris against fluconazole and AMB by gradient-MIC-strip (Etest) and broth microdilution-based MICRONAUT-AM-EUCAST (MCN-AM) assays. Clinical C. auris isolates (n = 121) identified by phenotypic and molecular methods were tested. Essential agreement (EA, ±1 two-fold dilution) between the two methods and categorical agreement (CA) based on the Centers for Disease Control and Prevention’s (CDC’s) tentative resistance breakpoints were determined. Fluconazole resistance-associated mutations were detected by PCR-sequencing of ERG11. All isolates identified as C. auris belonged to South Asian clade I and contained the ERG11 Y132F or K143R mutation. The Etest–MCN-AM EA was poor (33%) for fluconazole and moderate (76%) for AMB. The CA for fluconazole was higher (94.2%, 7 discrepancies) than for AMB (91.7%, 10 discrepancies). Discrepancies were reduced when an MCN-AM upper-limit value of 4 µg/mL for fluconazole-susceptible C. auris and an Etest upper-limit value of 8 µg/mL for the wild type for AMB were used. Our data show that resistance to fluconazole was underestimated by MCN-AM, while resistance to AMB was overestimated by Etest when using the CDC’s tentative resistance breakpoints of ≥32 µg/mL for fluconazole and ≥2 µg/mL for AMB. Method-specific resistance breakpoints should be devised for accurate AST of clinical C. auris isolates for proper patient management.","PeriodicalId":8151,"journal":{"name":"Antibiotics","volume":"25 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2024-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Evaluation of Etest and MICRONAUT-AM Assay for Antifungal Susceptibility Testing of Candida auris: Underestimation of Fluconazole Resistance by MICRONAUT-AM and Overestimation of Amphotericin B Resistance by Etest\",\"authors\":\"Mohammad Asadzadeh, Suhail Ahmad, Wadha Alfouzan, Inaam Al-Obaid, Bram Spruijtenburg, Eelco F. J. Meijer, Jacques F. 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引用次数: 0
摘要
耐多药念珠菌最近在医疗机构中引起了大规模爆发。对念珠菌进行快速、准确的抗真菌药敏试验(AST)对于正确处理侵袭性感染至关重要。商用 Sensititre Yeast One 和 Vitek 2 方法会低估或高估真菌对氟康唑和两性霉素 B (AMB) 的耐药性。本研究通过梯度-MIC-条带(Etest)和基于肉汤微量稀释的 MICRONAUT-AM-EUCAST (MCN-AM)检测法,评估了球孢子菌对氟康唑和两性霉素 B(AMB)的 AST 检测结果。对通过表型和分子方法鉴定的临床阴道杆菌分离株(n = 121)进行了检测。确定了两种方法之间的基本一致(EA,±1 倍稀释)和基于美国疾病控制和预防中心(CDC)暂定耐药性断点的分类一致(CA)。通过对 ERG11 进行 PCR 测序,检测出与氟康唑耐药性相关的突变。所有被鉴定为阿氏杆菌的分离物都属于南亚支系 I,并含有 ERG11 Y132F 或 K143R 突变。对于氟康唑,Etest-MCN-AM 的 EA 较差(33%),对于 AMB,EA 中等(76%)。氟康唑的 CA 值(94.2%,7 个差异)高于 AMB(91.7%,10 个差异)。当使用对氟康唑敏感的阿米巴痢疾杆菌的 MCN-AM 上限为 4 µg/mL 和对 AMB 的野生型的 Etest 上限为 8 µg/mL 时,差异有所减少。我们的数据显示,当使用疾控中心暂定的耐药性断点(氟康唑≥32 µg/mL,AMB≥2 µg/mL)时,MCN-AM 低估了对氟康唑的耐药性,而 Etest 则高估了对 AMB 的耐药性。应设计出针对特定方法的耐药性断点,以便对临床分离出的球孢子菌进行准确的 AST 检测,从而对患者进行适当的管理。
Evaluation of Etest and MICRONAUT-AM Assay for Antifungal Susceptibility Testing of Candida auris: Underestimation of Fluconazole Resistance by MICRONAUT-AM and Overestimation of Amphotericin B Resistance by Etest
Multidrug-resistant Candida auris has recently caused major outbreaks in healthcare facilities. Rapid and accurate antifungal susceptibility testing (AST) of C. auris is crucial for proper management of invasive infections. The Commercial Sensititre Yeast One and Vitek 2 methods underestimate or overestimate the resistance of C. auris to fluconazole and amphotericin B (AMB). This study evaluated the AST results of C. auris against fluconazole and AMB by gradient-MIC-strip (Etest) and broth microdilution-based MICRONAUT-AM-EUCAST (MCN-AM) assays. Clinical C. auris isolates (n = 121) identified by phenotypic and molecular methods were tested. Essential agreement (EA, ±1 two-fold dilution) between the two methods and categorical agreement (CA) based on the Centers for Disease Control and Prevention’s (CDC’s) tentative resistance breakpoints were determined. Fluconazole resistance-associated mutations were detected by PCR-sequencing of ERG11. All isolates identified as C. auris belonged to South Asian clade I and contained the ERG11 Y132F or K143R mutation. The Etest–MCN-AM EA was poor (33%) for fluconazole and moderate (76%) for AMB. The CA for fluconazole was higher (94.2%, 7 discrepancies) than for AMB (91.7%, 10 discrepancies). Discrepancies were reduced when an MCN-AM upper-limit value of 4 µg/mL for fluconazole-susceptible C. auris and an Etest upper-limit value of 8 µg/mL for the wild type for AMB were used. Our data show that resistance to fluconazole was underestimated by MCN-AM, while resistance to AMB was overestimated by Etest when using the CDC’s tentative resistance breakpoints of ≥32 µg/mL for fluconazole and ≥2 µg/mL for AMB. Method-specific resistance breakpoints should be devised for accurate AST of clinical C. auris isolates for proper patient management.