Rapid ganciclovir susceptibility assay using flow cytometry for human cytomegalovirus clinical isolates.
Rapid ganciclovir susceptibility assay using flow cytometry for human cytomegalovirus clinical isolates.
复制标题
使用流式细胞术对人巨细胞病毒临床分离株进行快速更昔洛韦敏感性测定。
DOI：
10.1128/aac.42.9.2326
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发表时间：
1998
影响因子：
4.9
通讯作者：
Crumpacker,CS
中科院分区：
文献类型：
--
作者：
McSharry,JJ;Lurain,NS;Drusano,GL;Landay,AL;Notka,M;O'Gorman,MR;Weinberg,A;Shapiro,HM;Reichelderfer,PS;Crumpacker,CS
采用荧光染料标记的抗人巨细胞病毒（HCMV）即刻早期抗原单克隆抗体和流式细胞术，对更昔洛韦（Ganciclovir）对HCMV临床分离株的敏感性进行了快速、定量和客观测定。用流式细胞仪分析了25株临床分离株的更昔洛韦药敏性，结果表明，更昔洛韦对其中19株临床分离株的半数抑制浓度（IC_（50））在1.14 ~ 6.66 μM之间，平均为4.32 μM（±1.93）（敏感; IC_（50）小于7 μM），2个分离株的IC_（50）分别为8.48和9.79 μM（部分耐药），4个分离株的IC_（50）大于96 μM（耐药）。通过空斑减少试验对这些临床分离株的药物敏感性进行比较分析，得出19株药物敏感分离株的IC 50小于6 μM，平均值为2.88 μM（±1.40），部分耐药分离株的IC 50为6 - 8 μM，4株耐药临床分离株的IC 50大于12 μM。通过流式细胞术测定获得的药物敏感和部分耐药临床分离株的IC 50与空斑减少测定获得的IC 50的比较显示可接受的相关性（r2= 0.473;P= 0.001），表明流式细胞术测定可替代劳动密集型、主观性和耗时的空斑减少测定。
Rapid, quantitative, and objective determination of the susceptibilities of human cytomegalovirus (HCMV) clinical isolates to ganciclovir has been assessed by an assay that uses a fluorochrome-labeled monoclonal antibody to an HCMV immediate-early antigen and flow cytometry. Analysis of the ganciclovir susceptibilities of 25 phenotypically characterized clinical isolates by flow cytometry demonstrated that the 50% inhibitory concentrations (IC50s) of ganciclovir for 19 of the isolates were between 1.14 and 6.66 μM, with a mean of 4.32 μM (±1.93) (sensitive; IC50less than 7 μM), the IC50s for 2 isolates were 8.48 and 9.79 μM (partially resistant), and the IC50s for 4 isolates were greater than 96 μM (resistant). Comparative analysis of the drug susceptibilities of these clinical isolates by the plaque reduction assay gave IC50s of less than 6 μM, with a mean of 2.88 μM (±1.40) for the 19 drug-sensitive isolates, IC50s of 6 to 8 μM for the partially resistant isolates, and IC50s of greater than 12 μM for the four resistant clinical isolates. Comparison of the IC50s for the drug-susceptible and partially resistant clinical isolates obtained by the flow cytometry assay with the IC50s obtained by the plaque reduction assay showed an acceptable correlation (r2= 0.473;P= 0.001), suggesting that the flow cytometry assay could substitute for the more labor-intensive, subjective, and time-consuming plaque reduction assay.