Flow cytometric determination of ganciclovir susceptibilities of human cytomegalovirus clinical isolates.
Flow cytometric determination of ganciclovir susceptibilities of human cytomegalovirus clinical isolates.
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流式细胞术测定人巨细胞病毒临床分离株的更昔洛韦敏感性。
DOI：
10.1128/jcm.36.4.958-964.1998
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发表时间：
1998
影响因子：
9.4
通讯作者：
Crumpacker,C
中科院分区：
文献类型：
--
作者：
McSharry,JM;Lurain,NS;Drusano,GL;Landay,A;Manischewitz,J;Nokta,M;O'Gorman,M;Shapiro,HM;Weinberg,A;Reichelderfer,P;Crumpacker,C
建立了人巨细胞病毒(HCMV)实验室分离株和临床分离株对更昔洛韦敏感性的流式细胞仪检测方法。该方法用荧光标记的抗人巨细胞病毒即刻、早期和晚期抗原的单抗鉴定巨细胞病毒感染的细胞，用流式细胞术检测和定量抗原阳性细胞的数量。用该方法测定了更昔洛韦对人巨细胞病毒AD169株的半数抑制浓度(IC50)为1.7μM，90%抑制浓度(IC90)为9.2μM，对更昔洛韦耐药株D6/3/1的IC50>12μM。更昔洛韦对药物敏感的临床分离株的IC50平均值为3.79μM(±2.6 0)。空斑减少法对这些临床分离株的平均IC50为2.80μM(±1.46)。将流式细胞仪分析结果与斑块减少分析结果进行比较，显示出可接受的偏差和精密度。对感染更昔洛韦耐药临床分离株的细胞进行的流式细胞仪和空斑减少分析未能显示晚期抗原阳性细胞或pFU的百分比减少，即使在96μ的更昔洛韦时也是如此。流式细胞术检测更昔洛韦对HCMV的敏感性是定量的、客观的、有可能自动化的，其结果在不同实验室之间是可重复性的。
A flow cytometric assay has been developed for the measurement of susceptibilities to ganciclovir of laboratory strains and clinical isolates of human cytomegalovirus (HCMV). The assay uses fluorochrome-labeled monoclonal antibodies to HCMV immediate-early and late antigens to identify HCMV-infected cells and flow cytometry to detect and quantitate the number of antigen-positive cells. By this assay, the 50 and 90% inhibitory concentrations (IC50and IC90, respectively) of ganciclovir for the AD169 strain of HCMV were 1.7 and 9.2 μM, respectively, and the IC50for the ganciclovir-resistant D6/3/1 derivative of the AD169 strain was greater than 12 μM. The ganciclovir susceptibilities of 17 HCMV clinical isolates were also determined by flow cytometric analysis of the effect of ganciclovir on late-antigen synthesis in HCMV-infected cells. The average IC50of ganciclovir for drug-sensitive HCMV clinical isolates was 3.79 μM (±2.60). The plaque-reduction assay for these clinical isolates yielded an average IC50of 2.80 μM (±1.46). Comparison of the results of the flow cytometry assays with those obtained from the plaque-reduction assays demonstrated acceptable bias and precision. Flow cytometric and plaque-reduction analysis of cells infected with ganciclovir-resistant clinical isolates failed to show a reduction in the percentage of late-antigen-positive cells or PFU, even at 96 μM ganciclovir. The flow cytometric assay for determining ganciclovir susceptibility of HCMV is quantitative, and objective, and potentially automatable, and its results are reproducible among laboratories.