[1]朱琳,尹立红,浦跃朴,等.荧光原位杂交法检测环境硝化细菌实验条件优化及应用[J].东南大学学报(自然科学版),2005,35(2):266-270.[doi:10.3969/j.issn.1001-0505.2005.02.023]
 Zhu Lin,Yin Lihong,Pu Yuepu,et al.Optimization and application of fluorescence in situ hybridization assay for detecting nitrifying bacteria in environmental samples[J].Journal of Southeast University (Natural Science Edition),2005,35(2):266-270.[doi:10.3969/j.issn.1001-0505.2005.02.023]
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荧光原位杂交法检测环境硝化细菌实验条件优化及应用()
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《东南大学学报(自然科学版)》[ISSN:1001-0505/CN:32-1178/N]

卷:
35
期数:
2005年第2期
页码:
266-270
栏目:
环境科学与工程
出版日期:
2005-03-20

文章信息/Info

Title:
Optimization and application of fluorescence in situ hybridization assay for detecting nitrifying bacteria in environmental samples
作者:
朱琳1 尹立红1 浦跃朴1 吕锡武2 李先宁2
1 东南大学公共卫生学院, 南京 210009; 2 东南大学土木工程学院, 南京 210096
Author(s):
Zhu Lin1 Yin Lihong1 Pu Yuepu1 Lü Xiwu2 Li Xianning2
1 School of Public Health, Southeast University, Nanjing 210009, China
2 College of Civil Engineering, Southeast University, Nanjing 210096, China
关键词:
荧光原位杂交法 硝化细菌 亚硝化细菌 硝酸菌
Keywords:
fluorescence in situ hybridization nitrifying bacteria nitrosobacteria nitrobacteria
分类号:
X172
DOI:
10.3969/j.issn.1001-0505.2005.02.023
摘要:
通过正交试验筛检荧光原位杂交(FISH)技术在检测环境中硝化细菌时的最佳及适合的实验条件.结果显示,样品预处理较优的条件为:热固定2 h,乙醇脱水3 min; FISH技术检测环境样本中亚硝化细菌的最佳实验条件为:杂交温度46 ℃,杂交时间3 h,洗脱液中NaCl浓度20 mmol/L; 检测硝酸菌最佳实验条件为:杂交温度46 ℃,杂交时间5 h,洗脱液中NaCl浓度60 mmol/L.应用该方案对太湖、玄武湖、南京锁金村污水处理厂活性污泥中硝化细菌分布进行分析,结果显示,环境样本中亚硝化细菌和硝酸菌含量分别是:太湖1.68×103, 0.54×103cells/mL; 玄武湖1.34×103, 0.38×103cells/mL; 锁金村污水处理厂活性污泥8.30×106, 0.52×106cells/g.FISH技术检测环境样品中硝化细菌与传统检测方法相比具有快速、简便、准确的优点,在研究环境微生物方面有较好的应用前景.
Abstract:
Experimental conditions of fluorescence in situ hybridization(FISH)for detecting nitrifying bacteria in environmental sample is optimized through orthogonal test. Results show that the optimal conditions for sample preparation are heat-fixing time of 2 h and ethanol-dehydration time of 3 min. The optimized conditions for detecting nitrosobacteria include a hybridized temperature of 46 ℃, a hybridized time of 3 h, and concentration of 20 mmol/L NaCl in hybridized solution. While for detecting nitrobacteria the conditions are a hybridized temperature of 46 ℃, a hybridized time of 5 h, and concentration of 60 mmol/L NaCl in hybridized solution. An effective procedure was established and used to detect nitrifying bacteria in Taihu Lake, Xuanwu Lake and activated sludge of the Suojincun sewage treatment plant in Nanjing. The detected contents of nitrobacteria and nitrosobacteria are respectively as follows: 1.68×103, 0.54×103cells/mL in Taihu; 1.34×103, 0.38×103cells/mL in Xuanwu Lake; 8.30×106, 0.52×106cells/g in activated sludge. Compared with traditional culture method, FISH assay is faster and more accurate. Therefore it has good application prospect in detection of environmental microorganisms.

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备注/Memo

备注/Memo:
基金项目: 国家高技术研究发展计划(863计划)资助项目(2002AA601011).
作者简介: 朱琳(1975—),女,硕士生; 尹立红(联系人),女,博士,教授,博士生导师, lhyin@seu.edu.cn.
更新日期/Last Update: 2005-03-20