Integrated on-chip microfluidic system with surface plasmon resonance biosensor for time-effective detection of legionella pneumophila in contaminated water

军团菌属革兰氏阴性细菌，常见于天然水生环境，特别是饮用水、冷却塔和热交换器系统。当易感宿主吸入雾化水或吸入含有细菌的水时，这些细菌会引起人类呼吸道疾病。军团菌攻击肺部并引起危及生命的肺炎。最近，与军团病的相关性变得越来越重要，据报道，魁北克爆发了180例病例和3例死亡。有50种细菌菌株属于军团菌，其中几乎一半是记录在案的人类病原体。细胞培养是检测水样中军团菌的常用方法。然而，这需要2天;当存在不止一种菌株时，甚至需要2周才能获得确定的结果。对于军团菌的种特异性检测，来自16 S rRNA的寡核苷酸通常用于聚合酶链反应以获得定性信息。最近，在使用芯片上实验室设备沿着用于检测病原体的传感技术方面投入了大量的努力。大多数可用的军团菌传感器专注于检测一种细菌菌株，当样品含有抑制军团菌生长的微生物时缺乏灵敏度。在这里，我们提出了一个实验室上的芯片设备的寡核苷酸来源于16 S rRNA检测军团菌的水质控制，通过集成的数字微流体无标记表面等离子体共振生物传感器。将实施适当的信号放大策略和优化的表面化学，以实现高灵敏度、特异性和缩短的测定时间。我们的方法需要几个原创性的方法，主要是开发一个适当的表面功能化的DNA探针的固定和光谱角SPR生物传感器工作在可见光谱与我们的兼容，

Legionella are gram-negative bacteria found in natural aquatic habitats, especially in potable water, cooling tower and heat exchanger systems. These bacteria cause respiratory disease in humans when a susceptible host inhales aerosolized water or aspirates water containing the bacteria. The legionella attack the lungs and cause a life threatening pneumonia. The relevance to legionnaire's has become ever more relevant as of late, with a reported outbreak in Quebec responsible for 180 cases and 3 deaths. There are 50 bacterial strains belonging to legionella, almost half of which are documented human pathogens. Cell cultures are a common method of testing legionella in water samples. However this requires 2 days; even 2 weeks when more than one strain is present, to obtain definitive results. For species-specific detections of Legionella, oligonucleotides derived from the 16S rRNA are often used in polymerase chain reaction to obtain qualitative information. More recently, there have been extensive efforts invested in the use lab-on-a-chip devices along with the sensing technology for the detection of pathogens. Most available sensors for legionella focus on detecting one strain of the bacteria and lacks sensitivity when samples contain microroganisms that inhibit legionella growth. Herein we propose a lab-on-a chip device for the oligonucleotides derived from the 16S rRNA detection of legionella for water quality control through the integration of a digital microfluidics within label free surface plasmon resonance biosensors. Adequate signal amplification strategies and optimized surface chemistry will be implemented to achieve high sensitivity, specificity and reduced assay time. Our methodology entails several original approaches, mainly the development of a proper surface functionalization for the immobilization of DNA probe and a spectro-angular SPR biosenor working in the visible spectrum compatible with our