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

军团菌是一种革兰氏阴性细菌，存在于自然水生环境中，特别是在饮用水、冷却塔和热交换器系统中。当易感宿主吸入雾化水或吸入含有细菌的水时，这些细菌会导致人类呼吸道疾病。军团菌攻击肺部并导致危及生命的肺炎。最近，与退伍军人症的相关性变得更加重要，据报道，魁北克爆发疫情导致180例病例和3人死亡。有50种细菌属于军团菌，其中近一半是已知的人类病原体。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；甚至需要2周(如果存在不止一种菌株)才能获得确定的结果。对于军团菌的物种特异性检测，从16S rRNA衍生的寡核苷酸经常用于聚合酶链式反应以获得定性信息。最近，在使用芯片实验室设备以及用于检测病原体的传感技术方面投入了广泛的努力。大多数现有的军团菌传感器专注于检测一种细菌菌株，当样本中含有抑制军团菌生长的微量细菌时，缺乏敏感性。在此，我们提出了一种芯片实验室装置，通过将数字微流控技术集成到无标记表面等离子体共振生物传感器中，用于检测军团菌16S rRNA的寡核苷酸，用于水质控制。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、特异度和缩短分析时间。我们的方法需要几种独创的方法，主要是开发一种适当的表面功能化来固定dna探针，以及一种工作在与我们的

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