Abstract: A set of primers and probes for simultaneous detection of Cymbidium mosaic virus (CymMV), Odontoglossum ringspot virus (ORSV), and Cymbidium ringspot virus (CymRSV) and a method for detecting CymMV, ORSV, and CymRSV, along with a method for their detection, are disclosed. The method involves designing multiplex real-time quantitative PCR detection primers and probes for CymMV, ORSV, and CymRSV and applying these primers and probes to the real-time quantitative PCR simultaneous detection of CymMV, ORSV, and CymRSV. It allows for faster detection of CymMV, ORSV, and CymRSV, taking only one-third of the time compared to uniplex real-time quantitative PCR technology, thereby reducing testing costs by approximately ? to ½ for each sample. The primers and probes are highly specific and sensitive, with a sensitivity as low as 1 to 10 copies. It provides an efficient and feasible detection method for early detection and prevention of CymMV, ORSV, and CymRSV.

Abstract: A method for improving a bio-coupling efficiency between a protein and a nucleic acid based on an ?-helix handle includes the following steps. First, the handle carrying the non-natural amino acid (H-tag) is designed. Then, a recombinant expression plasmid encoding a fusion protein containing the H-tag and the protein to be tested is constructed. Subsequently, the fusion recombinant protein containing the non-natural amino acid in the H-tag is expressed and purified. Finally, the non-natural amino acid in the H-tag-fused protein and the coupling group on the nucleic acid substrate are efficiently connected by click chemistry. Thea-helix handle is used to provide a controllable reaction condition on the protein surface for the non-natural amino acid, avoiding the complex structure, charge and polar nanoenvironment around the surface of the protein to be tested.

Abstract: A method for improving a bio-coupling efficiency between a protein and a nucleic acid based on an ?-helix handle includes the following steps. First, the handle carrying the non-natural amino acid (H-tag) is designed. Then, a recombinant expression plasmid encoding a fusion protein containing the H-tag and the protein to be tested is constructed. Subsequently, the fusion recombinant protein containing the non-natural amino acid in the H-tag is expressed and purified. Finally, the non-natural amino acid in the H-tag-fused protein and the coupling group on the nucleic acid substrate are efficiently connected by click chemistry. Thea-helix handle is used to provide a controllable reaction condition on the protein surface for the non-natural amino acid, avoiding the complex structure, charge and polar nanoenvironment around the surface of the protein to be tested.