Modifications of antifungal sensibility testing as suggested by CLSI document M27-A4: proposal for using different culture medium and buffer

协议(科学) 碳酸氢钠 氟康唑 特里斯 抗真菌 化学 色谱法 医学 生物 生物化学 微生物学 病理 有机化学 替代医学
作者
Edinaira Sulany Oliveira de Sousa,Silviane Bezerra Pinheiro,Ana Cláudia Alves Cortez,Kátia Santana Cruz,Érica Simplício de Souza,Márcia de Souza Carvalho Melhem,Hagen Frickmann,João Vicente Braga de Souza
出处
期刊:Diagnostic Microbiology and Infectious Disease [Elsevier BV]
卷期号:101 (3): 115488-115488 被引量:1
标识
DOI:10.1016/j.diagmicrobio.2021.115488
摘要

A common strategy in antifungal susceptibility testing is the utilization of the standardized protocol based on the microbroth dilution assay approach as described by the Clinical Laboratory Standards Institute (CLSI) (M27-A4). One major problem for laboratories in resource-limited countries with this protocol arises from the use of expensive culture media like RPMI-1640 and 3-N-morpholinopropanesulfonic acid (MOPS) buffer. One approach of circumventing this problem in cases of economic need is the evaluation of alternative culture media and buffers. The overall goal of this work was to investigate the influence of modifications in the protocol M27-A4 on diagnostic reliability. We performed univariate analyses evaluating (1) 2 different culture media (YNB and modified SAB); (2) three different buffers (sodium bicarbonate, Tris-HCL, and phosphate), as well as the influence of inoculum concentration (10 2 , 10 3 , 10 4 , 10 5 cells/mL), the influence of incubation time, and the influence of the assessment mode (visual, biological dye, and spectrophotometer). Our results suggested that (1) RPMI-1640 may be substituted by modified SAB and (2) MOPS buffer may be substituted by Tris-HCl buffer for defined analyses. By comparing the CLSI protocol and the alternative protocol proposed in the present study (modified SAB and Tris-HCl buffer) for the assessment of fluconazole susceptibility of eighteen yeasts (clinical isolates), similar results with both methodologies were recorded. We feel that this study should stimulate a discussion on the feasibility and evolution of the M27-A4 protocol in order to include pragmatic alternatives for resource-limited settings.
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