HKU-Shenzhen Institute of Research and Innovation (HKU-SIRI) , Shenzhen , Guangdong 518057 , China.
Department of Mechanical Engineering , The University of Hong Kong , Pokfulam Road , Hong Kong SAR , China.
Anal Chem. 2018 Nov 20;90(22):13173-13177. doi: 10.1021/acs.analchem.8b03673. Epub 2018 Oct 24.
In this study, we develop a method to detect multiple DNAs of foodborne pathogens by encapsulating emulsion droplets for loop-mediated isothermal amplification (LAMP). In contrast to the traditional bulk-phase LAMP, which involves a labor-intensive mixing process, with our method, different primers are automatically mixed with DNA samples and LAMP buffers after picoinjection. By directly observing and analyzing the fluorescence intensity of the resultant droplets, one can detect DNA from different pathogens, with a detection limit 500 times lower than that obtained by bulk-phase LAMP. We further demonstrate the ability to quantify bacteria concentration by detecting bacterial DNA in practical samples, showing great potential in monitoring water resources and their contamination by pathogenic bacteria.
在这项研究中,我们开发了一种通过包裹乳液液滴进行环介导等温扩增(LAMP)来检测食源性致病菌多种 DNA 的方法。与传统的体相 LAMP 相比,该方法需要进行繁琐的混合过程,而我们的方法则通过皮升级注射后自动将不同的引物与 DNA 样品和 LAMP 缓冲液混合。通过直接观察和分析所得液滴的荧光强度,可以检测来自不同病原体的 DNA,其检测限比体相 LAMP 低 500 倍。我们进一步通过检测实际样品中的细菌 DNA 证明了定量细菌浓度的能力,在监测水资源及其被致病菌污染方面具有很大的潜力。