用于无需培养的细菌计数的安培型基因传感器。
Amperometric genosensor for culture independent bacterial count.
作者信息
Jiang Xingxing, Liu Shuping, Yang Minghui, Rasooly Avraham
机构信息
Key Laboratory of Hunan Province for Water Environment and Agriculture Product Safety, College of Chemistry and Chemical Engineering, Central South University, Changsha, China, 410083.
National Cancer Institute, National Institutes of Health, Rockville, Maryland 20850, United States.
出版信息
Sens Actuators B Chem. 2019 Nov 15;299. doi: 10.1016/j.snb.2019.126944. Epub 2019 Aug 8.
Bacterial plate count for general assessment of water quality requires lengthy bacterial culturing. We report here a new DNA induced current genosensor for culture independent total bacteria determination. Since the amount of bacterial DNA is correlated to the number of bacteria, the genosensor measures the amount of bacterial DNA to determine bacterial count. The approach relies on bacteria lysis to release DNA which can react with molybdate to form redox molybdophosphate and measured electrochemically. Analysis of and demonstrated that the DNA generated current is highly correlated with the level of bacteria lysis which was confirmed by spectrometric measurement. Culture independent measurement of bacterial load suggests limit of detection is 21.9 CFU/mL, with linear range from 3×10 to 3×10 CFU/mL and correlation coefficient of 0.992. For analysis, the detection limit is 25.1 CFU/mL with the same linear range. The use of electrochemical microbial DNA quantitation for culture independent bacterial count is a new approach, the genosensor measurement is rapid (within 1 h) and has potential use for analysis of broad-spectrum bacteria for various applications.
用于水质总体评估的细菌平板计数需要长时间的细菌培养。我们在此报告一种新型的DNA诱导电流基因传感器，用于无需培养即可测定总细菌数。由于细菌DNA的量与细菌数量相关，该基因传感器通过测量细菌DNA的量来确定细菌数量。该方法依赖于细菌裂解以释放DNA，DNA可与钼酸盐反应形成氧化还原型磷钼酸盐，并通过电化学方法进行测量。对[具体内容1]和[具体内容2]的分析表明，产生的DNA电流与细菌裂解水平高度相关，这一点通过光谱测量得到了证实。无需培养即可测量[具体细菌1]的载量，检测限为21.9 CFU/mL，线性范围为3×10至3×10 CFU/mL，相关系数为0.992。对于[具体细菌2]分析，检测限为25.1 CFU/mL，线性范围相同。使用电化学微生物DNA定量法进行无需培养的细菌计数是一种新方法，该基因传感器测量速度快（1小时内），具有用于各种应用的广谱细菌分析的潜在用途。
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