基于多种微生物的毒性检测MTOXPlate冻干板制备及优化  

Preparation and optimization of lyophilised MTOXPlate for toxicity testing based on eleven different microorganisms

作　　者：林奕云[1,2,3,4] 张国霞[2,3,4] 许玫英[2,3,4] 贺丽苹[1] 孙国萍[2,3,4] 

机构地区：[1]华南农业大学食品科学学院,广东广州510642 [2]广东省微生物研究所广东省菌种保藏与应用重点实验室,广东广州510070 [3]广东省微生物应用新技术公共实验室,广东广州510070 [4]广东省华南应用微生物重点实验室-省部共建国家重点实验室培育基地,广东广州510070

出　　处：《微生物学通报》2013年第6期1096-1107,共12页Microbiology China

基　　金：广东联合基金项目(No.U1133005);广东省产学研项目(No.2011A090100030);广东省自然科学基金研究团队项目(No.9351007002000001);国家水体污染控制与治理科技重大专项项目(No.2008ZX07211-007)

摘　　要：【目的】制备基于多种微生物的生物毒性检测MTOXPlate冻干板,从而简化操作程序,使其能更灵敏、方便、快速地进行生物毒性测试。【方法】采用真空冷冻干燥法将指示微生物固定在微孔板中。以存活率为指标,利用荧光分光光度法确定冻干保护剂的组成。通过分析毒性响应灵敏度对检测条件进行优化。【结果】MTOXPlate的冻干保护剂组成为8%海藻糖、2%葡萄糖、2%甘露醇、1%谷氨酸钠。在此组成下,11株菌株的平均存活率最高,可达89.41%。复活介质与测试样品同时加入,在缩短了测试时间的同时也提高了检测灵敏度。该冻干板具有良好的灵敏度和重现性,能较好地满足污染物生物毒性测试的需要。【结论】MTOXPlate冻干板可作为一种新型的生物毒性测试技术,在生态毒理检测领域推广应用。[Objective] In order to simplify the procedure and improve the sensitivity of bio- logical toxicity testing method, the lyophilised MTOXPlate based on 11 genetically diverse microorganisms was prepared. ]Methods] The tested microbial strains were fixed on the mi- croplate by the vacuum freeze-drying method. Fluorescence spectrophotometry was used to determine the effects of several cryoprotectants on the freeze dried strains with the survival rate as the selecting index. The test condition was optimized through the sensitivity of toxicity detection. [Results] When the protective agent contains 8% trehalose, 2% glucose, 2% man- nitol and 1% sodium glutamate, the mean cell survival rate can reach 89.41% for 11 microbial strains used in this study. The resurrection medium and test sample were added to each well of the pre-prepared 96-well microplate at the same time. So that, short testing time and optimal detection sensitivity could be obtained. This lyophilised MTOXPlate exhibits high sensitivity and good reproducibility, which is suitable for the toxicity assessment. [Conclusion] MTOX- Plate could be a new biological toxicity test technology, which can be applied to the field of ecological toxicity testing.

关 键 词：11种微生物 MTOXPlate微孔板 冷冻干燥 毒性检测 

分 类 号：X832[环境科学与工程—环境工程]