Molecular Approaches for Low-Cost Point-of-Care Pathogen Detection in Agriculture and Forestry

Abstract

Early detection of plant diseases is a crucial factor to prevent or limit the spread of a rising infection that could cause significant economic loss. Detection test on plant diseases in the laboratory can be laborious, time consuming, expensive, and normally requires specific technical expertise. Moreover, in the developing countries, it is often difficult to find laboratories equipped for this kind of analysis. Therefore, in the past years, a high effort has been made for the development of fast, specific, sensitive, and cost-effective tests that can be successfully used in plant pathology directly in the field by low-specialized personnel using minimal equipment. Nucleic acid-based methods have proven to be a good choice for the development of detection tools in several fields, such as human/animal health, food safety, and water analysis, and their application in plant pathogen detection is becoming more and more common. In the present review, the more recent nucleic acid-based protocols for point-of-care (POC) plant pathogen detection and identification are described and analyzed. All these methods have a high potential for early detection of destructive diseases in agriculture and forestry, they should help make molecular detection for plant pathogens accessible to anyone, anywhere, and at any time. We do not suggest that on-site methods should replace lab testing completely, which remains crucial for more complex researches, such as identification and classification of new pathogens or the study of plant defense mechanisms. Instead, POC analysis can provide a useful, fast, and efficient preliminary on-site screening that is crucial in the struggle against plant pathogens.

Loop-Mediated Isothermal Amplification for Detection of Plant Pathogens in Wheat (Triticum aestivum)

“…The convenience of LAMP for detection of plant pathogens in wheat should be compared with other isothermal amplification methods. For instance, RPA does not require an initial heating step for DNA denaturation ( Baldi and La Porta, 2020 ). One of the main advantages of RPA is the lower temperature required to conduct the reaction, which is an improvement for field applications where access to electricity may be limited ( Panno et al, 2020 ).…”

“…However, important limitations of ELISA are low availability of enzyme-conjugated antibodies and low specificity ( Tomlinson and Boonham, 2008 ; Khater et al, 2017 ). Additionally, the production of monoclonal antibodies may be expensive ( Baldi and La Porta, 2020 ).…”

Noninvasive Technologies for Primate Conservation in the 21st Century

“…Another limitation of the MinION is the error rate, which some argue limits the utility for metabarcoding (Loit et al 2019), although there is a growing expectation that this is soon to improve, with rapid developments in flow cell chemistry and bioinformatics pipelines (Baldi and La Porta 2020;Chang et al 2020;Krehenwinkel 2019b). Currently field implementation of metabarcoding requires more testing and validation.…”

Nucleic Acid Extraction Methods and Technologies for On-Site Plant–Pathogen Diagnostics

“…The group of PCR-based detection methods is highly specific, sensitive, and reproducible; mostly known as the gold standard. This group also includes quantitative PCR (qPCR), nested PCR, multiplex PCR, and digital droplet PCR (ddPCR). − However, these approaches are expensive and labor- and time-consuming. They require dedicated equipment, trained personnel, and a high purity of nucleic acid.…”