A Rapid and Sensitive Loop-Mediated Isothermal Amplification (LAMP) Diagnostic Assay for Globodera rostochiensis

Abstract

Globodera rostochiensis, commonly known as the golden cyst nematode, is one of the most economically important quarantine pests of potato (Solanum tuberosum L.) worldwide. Its high reproductive rate and long-term persistence in soil make it difficult to manage, resulting in severe yield and quality losses in potato-growing regions. Rapid and accurate detection of G. rostochiensis is therefore critical for timely implementation of management strategies. In the present study, a loop-mediated isothermal amplification (LAMP) assay was developed for the rapid and accurate detection of G. rostochiensis by targeting the expansin B2 (expB2) gene. The assay successfully detected G. rostochiensis within 60 min, making it a rapid alternative to conventional molecular diagnostic approaches. The assay showed high specificity, with no cross-amplification with closely related cyst nematodes, G. pallida and Heterodera avenae. Furthermore, the assay exhibited high sensitivity, detecting genomic DNA concentrations as low as 1 pg/µl, whereas conventional PCR could detect only up to 100 pg/µl. Importantly, the assay was successfully validated for direct detection of G. rostochiensis from both artificially and naturally infested soil, showing its practical applicability for field diagnostics and quarantine inspections. Overall, the developed LAMP assay provides a rapid, highly specific, sensitive, and cost-effective diagnostic tool for reliable detection of G. rostochiensis in field samples and planting materials.