FISH法检测生物膜中硝化细菌流程的建立及优化  

Establishment of Experimental Conditions and Optimization of Fluorescence in situ Hybridization Assay for Detecting Nitrifying Bacteria in Biofilm

作　　者：张勇[1] 宋吟玲[1] 史俊[1] 陈宇[1] 

机构地区：[1]苏州科技学院环境科学与工程学院,江苏苏州215011

出　　处：《环境科技》2009年第4期34-37,共4页Environmental Science and Technology

摘　　要：通过试验筛检荧光原位杂交(FISH)技术在检测反应器生物膜中硝化细菌时适合的实验条件。结果显示,FISH技术检测生物膜中硝化细菌时,较好的预处理条件和杂交条件为:样品在热固定前不需1×PBS清洗;检测生物膜中硝化细菌的实验条件:①硝酸细菌:杂交温度46℃,杂交时间5h,洗脱液中c(NaCl)为55mmol/L;②亚硝酸细菌:杂交温度46℃,杂交时间3h,洗脱液中c(NaCl)为30～40mmol/L。Experimental conditions of fluorescence in situ hybridization （FISH） for detecting nitrifying bacteria in biotilm is optimized through test. Results show that the optimized pretreatment and hybridization conditions for detecting nitrifying bacteria include： Samples aren＇t cleaned by 1xPBS before heat fixed. Hybridization conditions of detecting nitrifying bacteria in biofilm are as follows： ①Nitrobacter： temperature is 46 ℃, hybridization time is 5 h, and sodium chloride concentration in the eluate is 55 mmol/L. ②Nitrosomonas： temperature is 46 ℃, hybridization time is 3h , and sodium chloride concentration in the eluate is 30 - 40 mmol/L.

关 键 词：硝化细菌 荧光原位杂交 杂交优化 

分 类 号：X2[环境科学与工程—环境科学]