自来水中细菌宏基因组DNA的浓度与细菌菌落总数的相关性分析  

The Correlation between Bacterial Metagenomic DNA Concentration and the Total Number of Bacteria Colonies in Drinking Water

作　　者：刘艳超[1] 郝文利[1] 宿庄[1] 高玉敏[1] 

机构地区：[1]内蒙古医科大学,呼和浩特010110

出　　处：《科学技术与工程》2015年第32期143-146,共4页Science Technology and Engineering

摘　　要：分析自来水中细菌宏基因组DNA浓度与细菌菌落总数之间的相关性,探索新的自来水细菌菌落总数检测方法。采用国标法检测自来水中的细菌菌落总数约为(165±5)CFU/m L,高于国标限值(100 CFU/m L)。通过计算得出,检测600 m L本实验水样中的细菌菌落总数相当于检测1 000 m L(1 L)100 CFU/m L的自来水。首先将600 m L及低于和高于该体积不同体积梯度的自来水经滤膜过滤收集菌体;然后使用细菌基因组提取试剂盒提取滤膜上的细菌宏基因组DNA,电泳分析提取的宏基因组质量;最后检测各个体积自来水宏基因组DNA浓度,根据检测结果分析自来水中宏基因组DNA含量与细菌菌落总数之间的相关性。提取的自来水宏基因组DNA主条带清晰,可以进行基因组DNA含量测定分析;自来水中细菌宏基因组DNA浓度与细菌菌落总数之间呈正相关的线性关系。建立一种通过检测自来水中细菌宏基因组DNA浓度来反映细菌菌落总数的检测方法,检测灵敏度可达7 CFU/m L,且省时、重复性好。Through analyzing the correlation between the bacterial metagenomic DNA concentration and the total count of bacterial colonies in drinking water, we explore a new method for the detection of total number of bacterial colonies in drinking water. The total number of bacteria colonies in the water from our lab was detected by a tradi- tional technology, and it was about （ 165 ±5） CFU/mL, which is higher than 100 CFU/mL that is the security boundary value which was made by Chinese health department in 2006. To limit the total number of bacterial colonies within 100 CFU/mL,the volume of the detection sample must below 600 mL. Firstly, the sample with different volumes higher or lower than 600 mL is treated with microfiltration; Secondly, extract the bacterial metagenomic DNA of filter membrane by Bacterial Genome Extraction Kit ; Thirdly, analyze the DNA extracted by agarose gel eleetrophoresis; Fourthly, determine the concentration of DNA extracted. Finally, analyze the correlation between the bacterial metagenomie DNA concentration and the total count of bacterial colonies in drinking water according to the experimental results. The bacterial metagenomic DNA extracted from different volumes of drinking water produce clear bands, and the DNA contents are enough for further analysis; the correlation between the bacterial metagenomic DNA concentration and the total count of bacterial colonies in drinking water is positive and linear. To establish a new method for detection of total number of bacterial colonies by analyzing the bacterial metagenomic DNA concentration in drinking water, the detection sensitivity can reach 7 CFU/mL, and the method is time-saving and with good repetitiveness.

关 键 词：宏基因组DNA 自来水 菌落总数 相关性 检测方法 

分 类 号：R123.1[医药卫生—环境卫生学]