Lee W, Kwak Y
Department of Laboratory Medicine, Ajou University School of Medicine, Suwon, Korea.
J Korean Med Sci. 1999 Feb;14(1):21-6. doi: 10.3346/jkms.1999.14.1.21.
The feasibility of flow cytometric antifungal susceptibility testing has been studied using the fluorescent anionic membrane potential probe, bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC4(3)]. The in vitro antifungal susceptibility testing of amphotericin B was performed on 8 Candida isolates from clinical specimens and 2 ATCC strains by flow cytometry with the results compared to those of the National Committee of Clinical Laboratory Standards (NCCLS) M27-T, broth macrodilution method. The flow cytometric method is based on an increase of fluorescence given out by DiBAC4(3) in fungi when they are killed by antifungal agents. Minimum inhibitory concentration (MIC) of amphotericin B ranged from 0.25 to 1 microg/mL. All results agreed within +/-2 dilution between the flow cytometric method and the M27-T method. MIC with ATCC strains were within recommended ranges of M27-T. The new flow cytometric method revealed a clear and distinct reproducible test end point. A four hr of incubation was sufficient for the test. In conclusion, flow cytometry using DiBAC4(3) is a rapid and accurate in vitro antifungal susceptibility testing method.
利用荧光阴离子膜电位探针双（1,3 - 二丁基巴比妥酸）三甲川草酚菁[DiBAC4(3)]研究了流式细胞术进行抗真菌药敏试验的可行性。采用流式细胞术对8株临床标本分离的念珠菌和2株美国典型培养物保藏中心（ATCC）菌株进行了两性霉素B的体外抗真菌药敏试验，并将结果与美国国家临床实验室标准委员会（NCCLS）M27 - T肉汤稀释法的结果进行比较。流式细胞术方法基于真菌被抗真菌药物杀死时DiBAC4(3)发出的荧光增加。两性霉素B的最低抑菌浓度（MIC）范围为0.25至1微克/毫升。流式细胞术方法与M27 - T方法的所有结果在±2倍稀释范围内一致。ATCC菌株的MIC在M27 - T推荐范围内。新的流式细胞术方法显示出清晰且可重复的检测终点。4小时的孵育时间足以完成试验。总之，使用DiBAC4(3)的流式细胞术是一种快速且准确的体外抗真菌药敏试验方法。
