采用美国临床和实验室标准协会（CLSI）方法及流式细胞术对新型隐球菌格鲁比变种AFLP1/VNI和加蒂隐球菌AFLP6/VGII进行两性霉素B的体外药敏试验。

In vitro susceptibility testing of amphotericin B for Cryptococcus neoformans variety grubii AFLP1/VNI and Cryptococcus gattii AFLP6/VGII by CLSI and flow cytometry.

作者信息

Morales Bernardina Penarrieta, Trilles Luciana, Bertho Álvaro Luiz, Junior Ivan Neves, de Oliveira Raquel de Vasconcellos Carvalhaes, Wanke Bodo, Lazéra Márcia dos Santos

机构信息

Laboratory Medical Mycology, National Institute of Infectious Diseases Evandro Chagas Oswaldo Cruz Foundation (INI-FIOCRUZ), Rio de Janeiro, Brazil.

出版信息

Mycoses. 2015 May;58(5):273-9. doi: 10.1111/myc.12310. Epub 2015 Mar 10.

DOI:10.1111/myc.12310

PMID:25756839

Abstract

Cryptococcus neoformans var. grubii AFLP1/VNI is the main causative agent of cryptococcosis associated with AIDS in the world. Cryptococcus gattii AFLP6/VGII causes mainly endemic primary infection in immunocompetent hosts. To determine the minimum inhibitory concentrations (MICs) of C. neoformans var. grubii AFLP1/VNI and C. gattii AFLP6/VGII against amphotericin B (AMB) in a short period of time, flow cytometry (FCM) with FUN-1 fluorochrome was used to compare with broth microdilution method (CLSI M27-A3). The minimum incubation period was evaluated by minimum fungicidal concentration procedure. Seventeen clinical isolates of C. neoformans var. grubii AFLP1/VNI and 18 of C. gattii AFLP6/VGII were analysed. The time for the determination of MICs by FCM was 2 h against 72 h by CLSI M27-A3 and the comparison of MIC showed a positive significant correlation (P = 0.048). It is important to highlight the role of the FCM as an alternative method to determine the MICs for AMB in within a day, with positive cost-benefit.

摘要

新型隐球菌格鲁比变种AFLP1/VNI是全球与艾滋病相关的隐球菌病的主要病原体。加蒂隐球菌AFLP6/VGII主要引起免疫功能正常宿主的地方性原发性感染。为了在短时间内确定新型隐球菌格鲁比变种AFLP1/VNI和加蒂隐球菌AFLP6/VGII对两性霉素B（AMB）的最低抑菌浓度（MIC），使用带有FUN-1荧光染料的流式细胞术（FCM）与肉汤微量稀释法（CLSI M27-A3）进行比较。通过最低杀菌浓度程序评估最短培养期。分析了17株新型隐球菌格鲁比变种AFLP1/VNI临床分离株和18株加蒂隐球菌AFLP6/VGII临床分离株。FCM测定MIC的时间为2小时，而CLSI M27-A3为72小时，MIC比较显示出显著正相关（P = 0.048）。重要的是要强调FCM作为一种替代方法在一天内确定AMB的MIC的作用，具有积极的成本效益。