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被误鉴定为哈氏假丝酵母菌的多重耐药耳念珠菌:通过基质辅助激光解吸电离飞行时间质谱和DNA测序进行特征分析以及其采用Vitek 2、CLSI肉汤微量稀释法和Etest法的抗真菌药敏谱变异性

Multidrug-Resistant Candida auris Misidentified as Candida haemulonii: Characterization by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry and DNA Sequencing and Its Antifungal Susceptibility Profile Variability by Vitek 2, CLSI Broth Microdilution, and Etest Method.

作者信息

Kathuria Shallu, Singh Pradeep K, Sharma Cheshta, Prakash Anupam, Masih Aradhana, Kumar Anil, Meis Jacques F, Chowdhary Anuradha

机构信息

Department of Medical Mycology, Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, India.

Department of Microbiology, Amrita Institute of Medical Sciences & Research Centre, Kochi, Kerala, India.

出版信息

J Clin Microbiol. 2015 Jun;53(6):1823-30. doi: 10.1128/JCM.00367-15. Epub 2015 Mar 25.

Abstract

Candida auris is a multidrug-resistant yeast that causes a wide spectrum of infections, especially in intensive care settings. We investigated C. auris prevalence among 102 clinical isolates previously identified as Candida haemulonii or Candida famata by the Vitek 2 system. Internal transcribed spacer region (ITS) sequencing confirmed 88.2% of the isolates as C. auris, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) easily separated all related species, viz., C. auris (n = 90), C. haemulonii (n = 6), C. haemulonii var. vulnera (n = 1), and Candida duobushaemulonii (n = 5). The in vitro antifungal susceptibility was determined using CLSI broth microdilution (CLSI-BMD), the Vitek 2 antifungal susceptibility test, and the Etest method. C. auris isolates revealed uniformly elevated fluconazole MICs (MIC50, 64 μg/ml), and an alarming percentage of isolates (37%) exhibited elevated caspofungin MICs by CLSI-BMD. Notably, 34% of C. auris isolates had coexisting elevated MICs (≥2 μg/ml) for both fluconazole and voriconazole, and 10% of the isolates had elevated coexisting MICs (≥2 μg/ml) to two additional azoles, i.e., posaconazole and isavuconazole. In contrast to reduced amphotericin B MICs by CLSI-BMD (MIC50, 1 μg/ml) for C. auris, elevated MICs were noted by Vitek 2 (MIC50, 8 μg/ml), which were statistically significant. Candida auris remains an unnoticed pathogen in routine microbiology laboratories, as 90% of the isolates characterized by commercial identification systems are misidentified as C. haemulonii. MALDI-TOF MS proved to be a more robust diagnostic technique for rapid identification of C. auris. Considering that misleading elevated MICs of amphotericin B by the Vitek AST-YS07 card may lead to the selection of inappropriate therapy, a cautionary approach is recommended for laboratories relying on commercial systems for identification and antifungal susceptibility testing of rare yeasts.

摘要

耳念珠菌是一种多重耐药酵母,可引起广泛的感染,尤其是在重症监护环境中。我们调查了102株先前经Vitek 2系统鉴定为哈氏假丝酵母或法塔假丝酵母的临床分离株中耳念珠菌的流行情况。内部转录间隔区(ITS)测序确认88.2%的分离株为耳念珠菌,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)轻松区分了所有相关菌种,即耳念珠菌(n = 90)、哈氏假丝酵母(n = 6)、哈氏假丝酵母脆弱变种(n = 1)和多布哈氏假丝酵母(n = 5)。使用CLSI肉汤微量稀释法(CLSI-BMD)、Vitek 2抗真菌药敏试验和Etest方法测定体外抗真菌药敏性。耳念珠菌分离株显示氟康唑MIC值普遍升高(MIC50,64μg/ml),并且通过CLSI-BMD法有相当比例(37%)的分离株显示卡泊芬净MIC值升高。值得注意的是,34%的耳念珠菌分离株同时存在氟康唑和伏立康唑MIC值升高(≥2μg/ml)的情况,10%的分离株对另外两种唑类药物即泊沙康唑和艾沙康唑的MIC值同时升高(≥2μg/ml)。与CLSI-BMD法测定的耳念珠菌两性霉素B MIC值降低(MIC50,1μg/ml)相反,Vitek 2测定的MIC值升高(MIC50,8μg/ml),具有统计学意义。耳念珠菌在常规微生物实验室中仍是一种未被注意的病原体,因为90%经商业鉴定系统鉴定的分离株被错误鉴定为哈氏假丝酵母。MALDI-TOF MS被证明是一种更可靠的诊断技术,可快速鉴定耳念珠菌。鉴于Vitek AST-YS07卡对两性霉素B的MIC值升高可能会导致选择不恰当的治疗方法,对于依赖商业系统进行罕见酵母菌鉴定和抗真菌药敏试验的实验室,建议采取谨慎的方法。

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