Detection of Ampelovirus and Nepovirus by Lab-on-a-Chip: A Promising Alternative to ELISA Test for Large Scale Health Screening of Grapevine

Abstract

The Ampelovirus Grapevine leafroll-associated virus 3 (GLRaV-3) and the Nepovirus Grapevine fanleaf virus (GFLV) are pathogens reported in many grapevine-growing areas all over the world, main causal agents of grapevine leafroll disease and grapevine fanleaf disease, respectively. Prevention of virus spread thanks to rapid diagnosis of infected plants is a key factor for control of both diseases. Although serological (e.g., enzyme-linked immunosorbent assay-ELISA test) and molecular methods are available to reveal the presence of the viruses, they turn out to be quite expensive, time-consuming and laborious, especially for large-scale health screening. Here we report the optimization of a lab-on-a-chip (LOC) for GLRaV-3 and GFLV detection, based on an electrochemical transduction and a microfluidic multichamber design for measurements in quadruplicate and simultaneous detection of both targets. The LOC detect GLRaV-3 and GFLV at dilution factors more than 15 times higher than ELISA, providing a higher sensitivity in the detection of both viruses. Furthermore, the platform offers several advantages as easy-to-use, rapid-test, portability and low costs, favoring its potential application for large-scale monitoring programs. Compared to other grapevine virus biosensors, our sensing platform is the first one to provide a dose-dependent calibration curve combined with a microfluidic module for sample analysis and a portable electronics providing an operator-independent read-out scheme.

Bioinspired Materials for Sensor and Clinical Applications: Two Case Studies

“…Aptamers and antibodies perform a conformational change that goes with an energy state transfer, and MIPs bind to ligands in their energy minimum state (or the fraction of the sample that is in that state), thus inducing no conformational changes. Lacking the adaptation, the dynamic of recognition is, in a certain sense, passive and works very well with inorganic ligands [31], offering the advantage of being accurate and providing highly selective results, as well as being fast and cheap, compared to conventional clinically applied methods for biomedical markers, pathogens, and toxin detection [35][36][37], such as enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) [36].…”

Multimodal Biosensing of Foodborne Pathogens

“…It exhibits the ability to sense GLRaV-3 and grapevine fanleaf virus (GFLV) at dilution factors over 15 times higher compared to ELISA, thereby offering enhanced sensitivity in virus identification ( Figure 5 B). This microfluidic platform is characterized by its simplicity, speed, miniaturization, and cost-effectiveness, making it compatible with large-scale monitoring evaluations [ 46 ]. Antonacci et al described an algal cytosensor designed for the electrochemical assessment of bacteria in wastewater using Chlamydomonas reinhardtii attached to carbon black (CB) nano-modified screen-printed microelectrodes.…”

Multimodal Biosensing of Foodborne Pathogens

“…This aptasensorʹs sensing and quantification limits were established to be 10 and 4.5 CFU/mL, respectively, with a linear range of approximately 101-108 CFU/mL [61]. Buja et al (2022) presented a microfluidics-based chip for ampelovirus and nepovirus detection. This chip features a multi-chamber design, enabling the simultaneous and rapid identification of these targets.…”