Kaminsky R, Brun R
Swiss Tropical Institute, Basel.
Acta Trop. 1993 Sep;54(3-4):279-89. doi: 10.1016/0001-706x(93)90100-p.
In vitro test methods have been developed both for determining the sensitivity of isolates of pathogenic African trypanosomes and for evaluating new compounds for antitrypanosomal activity. The principles of the assays and their main advantages or drawbacks are presented and discussed. In vitro assays which do not require trypanosomes pre-adapted to culture conditions are the Drug Incubation Infectivity Test (DIIT) and the [3H]Hypoxanthine Incorporation Assay. Chemosensitivity tests which do require continuous growth of trypanosomes in vitro include photometric, fluorescence, growth inhibition, long-term viability and metacyclic incubation assays. Evidence is presented to use metacyclic or bloodstream forms in such assays but to avoid procyclic trypanosomes. The drug sensitivity of a homogeneous trypanosome population can be quantified by using photometric, fluorescence or growth inhibition assays lasting 1-3 days. Small numbers of resistant organisms hidden in a sensitive population can be detected employing long-term viability assays (7-10 days). Final selection of the assay to be employed will depend on the parameter to be investigated, and the equipment available.
已经开发出体外测试方法,用于确定致病性非洲锥虫分离株的敏感性以及评估具有抗锥虫活性的新化合物。本文介绍并讨论了这些测定方法的原理及其主要优点或缺点。不需要将锥虫预先适应培养条件的体外测定方法有药物孵育感染性试验(DIIT)和[3H]次黄嘌呤掺入试验。确实需要锥虫在体外持续生长的化学敏感性试验包括光度法、荧光法、生长抑制法、长期活力法和循环后期孵育试验。有证据表明在这类试验中应使用循环后期或血流形式的锥虫,而应避免使用前循环期锥虫。通过持续1至3天的光度法、荧光法或生长抑制试验,可以对均匀锥虫群体的药物敏感性进行量化。采用长期活力试验(7至10天)可以检测出隐藏在敏感群体中的少量抗性生物体。最终选择采用何种试验将取决于要研究的参数以及可用的设备。