A transcription factor-based capture strategy for LAMP detection of wheat stripe mosaic virus.

Loop-mediated isothermal amplification (LAMP) is a well-established nucleic acid amplification method widely used for rapid and sensitive detection of plant viruses. Despite its robustness, downstream detection and multiplexing of LAMP products remain methodological challenges. Here, we describe a novel strategy for the capture and detection of LAMP amplification products based on the incorporation of transcription factor recognition sequences into LAMP primers. In this approach, a DNA-binding motif recognized by the transcription factor GCN4 was rationally introduced into the Forward Inner Primer (FIP), while the corresponding Backward Inner Primer (BIP) was biotinylated. Following standard LAMP amplification, the resulting products were specifically captured via GCN4-DNA interactions and detected using protein affinity-based assays, including ELISA and dot blot formats. Using wheat stripe mosaic virus (WhSMV) as a model target, we demonstrate that the incorporation of transcription factor binding sites into LAMP primers does not interfere with amplification and enables specific post-amplification capture and detection of LAMP products. Importantly, this study does not aim to revalidate LAMP performance or sensitivity, but rather to establish a proof-of-concept methodology for sequence-specific capture of LAMP amplicons. This platform provides a flexible methodological framework for the development of affinity-based detection and multiplexing strategies compatible with LAMP-generated products, expanding the toolkit available for nucleic acid detection workflows.