摘要
目的利用宏基因组技术建立一种新的自来水细菌总数检测方法。方法采用国标法检测自来水中的菌落总数约为(165±5)CFU/ml。如果检测样品量为1 L,折算1 L符合国标限值的自来水对应600 ml本实验用水,滤膜过滤法提取国标限值及其他体积该自来水的细菌总基因组,电泳分析,计算样品与限值总基因组主条带的亮度比值,比值近似或小于1则样品合格,否则不合格。结果借助Image J图像分析软件对总基因组电泳结果进行亮度分析,85、120、170与100 CFU/ml的自来水中的总基因组亮度比分别为0.95、1.6、1.9。结论宏基因组技术可以用于自来水中细菌总数的定量分析,该方法检测的结果与国标法一致,区分度约为15 CFU/ml,且相对省时、重复性好。
Objective To establish a new method to count the total number of bacterial colonies in tap water based on the metagenomic technology. Methods The total number of bacterial colonies in tap water collected from the lab and detected by the national standard method was about （165 ± 5） CFU/ml. According to the security boundary value of bacterial colonies in tap water, the metagenome [ （165 ±5） CFU/ml] in the 600 ml lab water was almost equal to 100 CFU/ml in the 1,000 ml tap wa- ter, which was made by Chinese health department in 2006. The metagenome in the 500, 600, 700 and 1,000 ml lab water was extracted respectively, and the first three were regarded as 1,000 ml 85, 100 and 120 CFU/ml tap water samples, successively. After that, 1% agarose gel electrophoresis was employed to detect the metagenomes. Then metagenomic concentrations of the four different volumes of the lab water were analyzed by Image J （an image analysis software）. Concentration ratios of the other 3 volumes of metagenome to that of the 600 ml were figured out. If the ratio was more than 1, the sample was not qualified. Results The genome concentration ratios were 0.95, 1.6 and 1.9 from 1,000 ml 85, 120 and 170 CFU/ml tap water com- pared with that of 100 CFU/ml. The results worked out by the metagenomic technology were consistent with those of traditional national standard method. Conclusions The metagenomic technology is suitable for quantitative estimation of the total num- ber of bacterial colonies in tap water. The results detected by this new method are not only consistent with those of the national standard method, but also has the advantages of saving time and repeatability, with the distinguishing index of 15 CFU/ml.
出处
《实用预防医学》
CAS
2016年第5期624-627,共4页
Practical Preventive Medicine
关键词
宏基因组技术
自来水
菌落总数
检测方法
Metagenomic technology
Tap water
Colony count
Detection method
作者简介
刘艳超（1983-），女，硕士研究生，助教，主要从事卫生微生物教学及科研工作。
通讯作者：高玉敏（1972-），女，教授，博士，主要从事分子流行病学研究，E-mail：gaoyumin0712@163．com.
