Simple and rapid detection of bacteria using a nuclease-responsive DNA probe†,Analyst

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Simple and rapid detection of bacteria using a nuclease-responsive DNA probe†
Analyst ( IF 3.6 ) Pub Date : 2017-11-24 00:00:00 , DOI: 10.1039/c7an01384a
Kyung Jin Lee _{1,

2,

3,

4,

5} , Wang Sik Lee _{2,

3,

4,

5,

6} , Ahreum Hwang _{2,

3,

4,

5,

7} , Jeong Moon _{2,

3,

4,

5} , Taejoon Kang _{1,

2,

3,

4,

5} , Kyoungsook Park _{1,

2,

3} , Jinyoung Jeong _{1,

2,

3,

4,

5}

Affiliation

We demonstrate simple and rapid bacterial detection using a nuclease-responsive DNA probe. The probe consisting of a fluorescent dye and a quencher at the 5′ and 3′ termini, respectively, was designed to be cleaved by nucleases such as endonucleases, exonucleases, and DNases, which are released from bacteria using an optimized lysis buffer. The fluorescence signal of the cleaved DNA probe correlates with the number of Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus, and the detection limit was 10³ CFU for E. coli and 10⁴ CFU for S. aureus. Moreover, this method is specific for live bacteria and takes just one minute to get the signal including sample collection. These features make the present bacterial detection method a powerful on-site bacterial contamination assay which is simple, rapid, and quantitative.

中文翻译：

使用核酸酶反应性DNA探针简单快速地检测细菌†

我们展示了使用核酸酶反应性DNA探针的简单，快速的细菌检测。探针分别由5'和3'末端的荧光染料和淬灭剂组成，被设计成可被核酸酶（例如核酸内切酶，核酸外切酶和DNase）切割，并使用优化的裂解缓冲液从细菌中释放出来。革兰氏阴性数目的切割的DNA探针相关因素的荧光信号大肠杆菌和革兰氏阳性金黄色葡萄球菌，检出限为10个³ CFU用于大肠杆菌和10个⁴ CFU为金黄色葡萄球菌。而且，这种方法是针对活细菌的，仅需一分钟即可获得包括样品采集在内的信号。这些特征使本细菌检测方法成为简单，快速和定量的强大的现场细菌污染测定。

更新日期：2017-11-24

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