A Listeria monocytogenes aptasensor on laser inscribed graphene for food safety monitoring in hydroponic water

Consumption of fresh produce, such as leafy greens, is often encouraged as part of a healthy diet. Hence, indoor facilities for hydroponic production of leafy greens are increasingly being established. However, fresh produce entails a higher risk of microbial foodborne illnesses than processed foods. Listeria monocytogenes is a major source of fresh produce contamination and is among the leading causes of severe foodborne illnesses in the United States, with a 16% mortality rate. Tools for rapid monitoring are needed for pathogens such as L. monocytogenes to prevent outbreaks. In this manuscript, we have demonstrated the feasibility of a multi-aptamer approach for development of label-free aptasensors targeting L. monocytogenes in irrigation water for lettuce hydroponic production. We use screening studies with surface plasmon resonance to rationally develop mixtures of relevant aptamers for targeting L. monocytogenes. Based on this screening, multiple aptamers targeting extracellular structures on intact L. monocytogenes were tethered to platinum-modified laser inscribed graphene electrodes. This is the first report of a L. monocytogenes biosensor based on laser inscribed graphene. We show that mixing multiple aptamers with varying affinity improves the diagnostic performance over one aptamer alone in complex sample matrices (lettuce hydroponic water). Multi-aptamer biosensors showed high accuracy for L. monocytogenes and were at least three times more selective than Escherichia coli (Crooks, K12, O157:H7) with an accuracy of 85%. The limit of detection (10 CFU/10 mL) is based on data which were significantly different after calibration toward L. monocytogenes or E. coli (Crooks) and validated against gold standard molecular analysis (polymerase chain reaction). Rapid screening of pathogens is a global need to meet food safety and water quality regulations. This study shows the importance of sensors targeting more than one bacterial surface structure in complex samples relevant to the food-water nexus.