Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.
Optimization of a Fluorescence-Based Assay for Large-Scale Drug Screening against Babesia and Theileria Parasites.
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DOI：
10.1371/journal.pone.0125276
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Igarashi I
中科院分区：
文献类型：
--
作者：
Rizk MA;El-Sayed SA;Terkawi MA;Youssef MA;El Said el Sel S;Elsayed G;El-Khodery S;El-Ashker M;Elsify A;Omar M;Salama A;Yokoyama N;Igarashi I
一个快速，准确的方法来评估抗巴贝虫药物在大规模的发现新的化疗药物对巴贝虫寄生虫。在本研究中，我们评估了基于荧光的测定法在体外培养中测定抗巴贝斯虫化合物对牛和马血液寄生虫的效力的有用性。使用三种不同的血细胞比容（HCT; 2.5%、5%和10%），每日不更换培养基。高通量筛选试验的结果显示，牛巴伯虫的最佳HCT为2.5%，马巴伯虫和泰勒虫的最佳HCT为5%。通过荧光法和显微镜法获得的乙酰氨基二氮烯的IC 50值无显著差异。同样地，毛地黄黄酮、咯萘啶四磷酸盐、印楝内酯、gedunin和依诺沙星的IC 50值在两种方法之间没有差异。总之，我们的基于荧光的测定使用低HCT，并且不需要每天更换培养基，使得其非常适合于针对感染牛和马的巴氏杆菌和泰勒虫寄生虫的体外大规模药物筛选。
A rapid and accurate assay for evaluating antibabesial drugs on a large scale is required for the discovery of novel chemotherapeutic agents against Babesia parasites. In the current study, we evaluated the usefulness of a fluorescence-based assay for determining the efficacies of antibabesial compounds against bovine and equine hemoparasites in in vitro cultures. Three different hematocrits (HCTs; 2.5%, 5%, and 10%) were used without daily replacement of the medium. The results of a high-throughput screening assay revealed that the best HCT was 2.5% for bovine Babesia parasites and 5% for equine Babesia and Theileria parasites. The IC50 values of diminazene aceturate obtained by fluorescence and microscopy did not differ significantly. Likewise, the IC50 values of luteolin, pyronaridine tetraphosphate, nimbolide, gedunin, and enoxacin did not differ between the two methods. In conclusion, our fluorescence-based assay uses low HCT and does not require daily replacement of culture medium, making it highly suitable for in vitro large-scale drug screening against Babesia and Theileria parasites that infect cattle and horses.
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影响因子：
1.5
作者：
Hötzel, I;Suarez, CE;Palmer, GH
通讯作者：
Palmer, GH
DOI：
10.4269/ajtmh.1991.44.11
发表时间：
1991-01-01
影响因子：
3.3
作者：
MAKLER, MT;RIES, LK;HINRICHS, DJ
通讯作者：
HINRICHS, DJ
影响因子：
4.9
作者：
DIVO, AA;SARTORELLI, AC;BIA, FJ
通讯作者：
BIA, FJ
DOI：
10.4269/ajtmh.1979.28.184
发表时间：
1979-01-01
影响因子：
3.3
作者：
RUEBUSH, TK;RUBIN, RH;SCHULTZ, MG
通讯作者：
SCHULTZ, MG
影响因子：
4.9
作者：
Bork, S;Yokoyama, N;Igarashi, I
通讯作者：
Igarashi, I