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Antibiotic and cytotoxic drug susceptibility assays using resazurin and poising agents

机译:使用刃天青和平衡剂的抗生素和细胞毒性药物敏感性测定

摘要

Growth inhibition assays for both microorganisms and mammalian cells are performed by incubating the microorganisms or cells in a growth medium in the presence of resazurin and a suspected growth inhibiting agent. By observing a color change accompanied by the reduction of resazurin to resorfurin, continued cell viability can be determined. The present invention reduces system errors by inhibiting the autoreduction of resazurin by the growth media by incorporating suitable redox stabilizing agents in the medium. In a preferred embodiment the poising agent is potassium ferrocyanide, ferric salt or ferricinium and the medium is maintained at a potential from about +0.3 volts to about +0.45 volts. A second poising agent can be used to inhibit the reduction of resorfin to dihydroresorfin. Preferred second poising agents are methylene blue, toluidine blue, azure I and gallocyanide in amounts to maintain the potential of the growth medium above -0.1 volts.
机译:通过在刃天青和可疑的生长抑制剂存在下将微生物或细胞在生长培养基中孵育来进行微生物和哺乳动物细胞的生长抑制测定。通过观察颜色变化,伴随着刃天青素还原为间苯二酚,可以确定持续的细胞活力。通过在培养基中掺入合适的氧化还原稳定剂,本发明通过抑制生长培养基中刃天青的自动还原来减少系统错误。在一个优选的实施方案中,所述起泡剂是亚铁氰化钾,铁盐或铁盐,并且所述介质保持在约+0.3伏至约+0.45伏的电势。可以使用第二种平衡剂来抑制间苯二酚还原为二氢间苯二酚。优选的第二平衡剂是亚甲基蓝,甲苯胺蓝,天青I和没食子氰化物,其数量应保持生长培养基的电势在-0.1伏以上。

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