PDF(Pubmed)
Abstract：
Legionella pneumophila has been pinpointed by the World Health Organization as the highest health burden of all waterborne pathogens in the European Union and is responsible for many disease outbreaks around the globe. Today, standard analysis methods (based on bacteria culturing onto agar plates) need several days (~12) in specialized analytical laboratories to yield results, not allowing for timely actions to prevent outbreaks. Over the last decades, great efforts have been made to develop more efficient waterborne pathogen diagnostics and faster analysis methods, requiring further advancement of microfluidics and sensors for simple, rapid, accurate, inexpensive, real-time, and on-site methods. Herein, a lab-on-a-chip device integrating sample preparation by accommodating bacteria capture, lysis, and DNA isothermal amplification with fast (less than 3 h) and highly sensitive, colorimetric end-point detection of L. pneumophila in water samples is presented, for use at the point of need. The method is based on the selective capture of viable bacteria on on-chip-immobilized and -lyophilized antibodies, lysis, the loop-mediated amplification (LAMP) of DNA, and end-point detection by a color change, observable by the naked eye and semiquantified by computational image analysis. Competitive advantages are demonstrated, such as low reagent consumption, portability and disposability, color change, storage at RT, and compliance with current legislation.
摘要：
肺炎军团菌已被世界卫生组织确定为欧盟所有水传播病原体中最高的健康负担，并且是全球许多疾病爆发的原因。今天,标准分析方法（基于细菌培养在琼脂平板上）需要几天（〜12）在专门的分析实验室产生的结果，不允许及时采取行动防止疫情爆发。在过去的几十年里，已经做出了巨大的努力来开发更有效的水传播病原体诊断和更快的分析方法，需要进一步改进微流体和传感器，快速,准确,便宜,实时,和现场方法。在这里，通过容纳细菌捕获来集成样品制备的芯片实验室装置，裂解，和DNA等温扩增快速（小于3小时）和高度敏感，提出了水样中嗜肺乳杆菌的比色终点检测，在需要的时候使用。该方法基于在芯片上固定和冻干的抗体上选择性捕获活细菌，裂解，DNA的环介导扩增（LAMP），通过颜色变化进行终点检测，肉眼可观察，并通过计算图像分析半量化。竞争优势得到证明，如低试剂消耗，便携性和一次性，颜色变化,储存在RT，并遵守现行法律。
