荧光原位杂交技术检测聚磷菌方法的研究  

Research of Detecting PAOs by Fluorescence in Situ Hybridization

作　　者：史俊[1] 宋吟玲[1] 

机构地区：[1]苏州科技学院环境科学与工程系,江苏苏州215011

出　　处：《环境科学与技术》2009年第B12期142-146,共5页Environmental Science & Technology

摘　　要：研究通过正交实验和平行实验选择对纯培养的聚磷菌进行荧光原位杂交检测的最佳方法。经试验确定，聚磷菌探针杂交时间为2．5h，杂交温度为46℃，甲酰胺浓度为40％。在利用2种或2种以上探针进行荧光原位杂交时，用重复多色法比同步多色法效果要好，即分别用探针与样品杂交的效果好于同时用探针与样品杂交的效果。在用重复双色FISH法时，当两种探针的杂交温度，杂交液甲酰胺浓度等条件相近而杂交时间差异较大时可以在一种探针杂交完成后不经过洗涤就立刻进行再次杂交；而当两种探针杂交条件差异较大时，最好是一种探针杂交完成后洗涤再进行再次杂交。利用确定的实验方法，可以利用不同探针对活性污泥中不同种细菌进行同时检测，获取更多的活性污泥微生物信息。In this study orthogonal experiments and parallel experiments was used to select the best way to detect pure PAOs by fluorescence in situ hybridization. The best temperature suitable for the PAO651 probe for hybridization is 46℃, the concentration of formamide for hybridization is 40%, hybridization time is 2.5 hours. When using two or more kinds of probes, the result of duplicate multieolor is better than synchronous multicolor, that is the result of hybridization which used separately probe is better than the result of hybridization which use probe at the same time. For duplieate twocolor FISH, when the hybridization temperature and formamide concentration in the hybridization solution of the two probes are similar but the hybridization time can be quite different, it can take a hybridization with a probe without washing then re-hybridization with another probe; and when the hybridization conditions of the two probes are very different, it is better to take a washing after the completion of a hybridization with a probe then re-hybridization with another probe. Use the method determined by experimental, we can use different probes to detect different kinds of activated sludge bacteria, to get more information of the activated sludge microorganisms.

关 键 词：荧光原位杂交 探针 同步双色 重复双色 

分 类 号：X703[环境科学与工程—环境工程]