Division of Biology, Food and Drug Administration, Food and Drug Administration, Center for Devices and Radiological Health, Office of Device Evaluation, Silver Spring, MD, USA.
Eye Contact Lens. 2012 Nov;38(6):363-7. doi: 10.1097/ICL.0b013e31826e16e4.
To optimize the growth, culture, and life cycle conditions for testing multipurpose solutions (MPS) against Acanthamoebatrophozoites and cysts to better inform the development of an appropriate test protocol.
Two strains of Acanthamoeba castellanii were grown using 2 different methods, bacterized and axenic. Amoebae grown from both methods and from both strains were treated with 4 different MPS as trophozoites and cysts, which were generated using 4 encystment methods. Experiments were run in triplicate with controls. A 5-tube most probable number method was used to enumerate the survivors and to determine the log kills. Statistical analysis was performed using effect screening.
There was a marked difference in effectiveness among solutions, which varied with growth conditions (P<0.0001) and encystment method. Growth medium affected survival. In addition, there was a significant difference in cyst survival, which was dependent on encystment method (P=0.0013). The strain used was less of a factor in trophozoite resistance to MPS. Cyst resistance to MPS varied depending on which strain was used, but it was not a significant factor.
When designing a contact lens solution efficacy testing protocol for Acanthamoeba, care should be taken to control for variables that may distort results. An appropriate protocol should include growing Acanthamoeba bacterized and allowing them to encyst naturally. By choosing optimized testing conditions, a more realistic efficacy of contact lens solutions can be determined which will result in better and safer products on the market.
优化多用途溶液（MPS）针对阿米巴滋养体和包囊的生长、培养和生命周期条件的测试，以更好地为适当的测试方案的制定提供信息。
使用两种不同的方法（细菌化和无菌）培养两种棘阿米巴属（Acanthamoeba）Castellanii 菌株。从这两种方法和这两种菌株生长的变形虫都用 4 种不同的 MPS 处理，这些 MPS 作为滋养体和包囊，使用 4 种包囊方法生成。实验一式三份进行，有对照。使用 5 管最可能数法（MPN）来计数幸存者并确定对数杀灭。使用效应筛选进行统计分析。
溶液的有效性存在显著差异，差异随生长条件（P<0.0001）和包囊方法而异。生长培养基影响存活。此外，囊的存活有显著差异，这取决于包囊方法（P=0.0013）。使用的菌株对 MPS 的滋养体抗性的影响较小。MPS 对囊的抗性取决于使用的菌株，但不是一个显著因素。
在设计棘阿米巴接触镜溶液功效测试方案时，应注意控制可能扭曲结果的变量。适当的方案应包括培养细菌化的棘阿米巴并允许它们自然包囊。通过选择优化的测试条件，可以确定更符合实际的接触镜溶液功效，从而在市场上提供更好、更安全的产品。
