Microfluidic device for the point of need detection of a pathogen infection biomarker in grapes

• Published in: Analyst (London) 2019-08, Vol.144 (16), p.4871-4879
• Main Authors: Brs, Eduardo J. S, Fortes, Ana Margarida, Chu, Virginia, Fernandes, Pedro, Conde, Joo Pedro
• Format: Article
• Language: English
• Subjects: Biomarkers; Biomarkers - analysis; Biomarkers - chemistry; Colorimetry; Colorimetry - methods; Crops; Dicarboxylic Acids - analysis; Dicarboxylic Acids - chemistry; Dilution; Enzyme Inhibitors - analysis; Enzyme Inhibitors - chemistry; Enzymes; Enzymes, Immobilized - chemistry; Fruit and Vegetable Juices - analysis; Grapes; Lab-On-A-Chip Devices; Limit of Detection; Microfluidic Analytical Techniques - methods; Microfluidic devices; Monitoring; Monophenol Monooxygenase - antagonists & inhibitors; Monophenol Monooxygenase - chemistry; Nanoparticles; Pathogens; Plant Diseases - microbiology; Thin films; Tyrosinase; Vitis - microbiology
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/c9an01002e

Bacterial, fungal and viral infections in plant systems are on the rise, most of which tend to spread quickly amongst crops. These pathogens are also gaining resistance to known treatments, which makes their early detection a priority to avoid extensive loss of crops and the spreading of disease to animal systems. In this work, we propose a microfluidic platform coupled with integrated thin-film silicon photosensors for the detection of pathogen infections in grapes. This detection was achieved by monitoring the concentration of Azelaic Acid (AzA). This small organic acid plays a significant role in the defense mechanism in plant systems. In this platform, the enzyme tyrosinase was immobilized on microbeads inside a microfluidic system. By colorimetric monitoring of the inhibitory effect of AzA on the enzyme tyrosinase in real time, it was possible, in under 10 minutes, to detect different concentrations of AzA in both buffer and spiked solutions of grape juice, in both cases with limits of detection in the 510 nM range. In addition, with this microfluidic device, it was possible to clearly distinguish infected from healthy grape samples at three different grape maturation points. Healthy grape samples showed AzA concentrations in the range of 1020 nM (post-dilution) while infected samples have an estimated increase of AzA of 1030, results which were confirmed using HPLC. In both juice and grape samples an integrated sample preparation stage that decreases the phenol content of the solutions was required to achieve fit-for-purpose sensitivities to AzA. Due to the rise of pathogenic infections amongst crops, there is an increased need for point-of-need monitoring of plant health. In this work we present a portable system capable of detecting signs of infection in grapes using a microfluidic device.