Transboundary and Emerging Diseases / 2024 / Article / Tab 1 / Research Article
Development of Rapid Isothermal Detection Methods Real-Time Fluorescence and Lateral Flow Reverse Transcription Recombinase-Aided Amplification Assay for Bovine Coronavirus Table 1 The primers and probes, sequences of real-time RT-RAA primers and exo probe, RT-RAA-LFD primers and nfo probe.
Methods Primers Sequences (5′→3′) Position of N (gene/bp) Real-time RT-RAA Exo probe ACCCAAGTAGCGATGAGGCTATTCCGAC[FAM-dT]A[THF]G[BHQ-dT]TTCCGCCTGGCACGG-C3Spacer 494–541 This study F1 TATGGCACCGATATTGACGGAGTCTTCTGG 412–441 This study R1 GAGCAGACCTTCCTGAGCCTTCAATATAGT 557–586 This study F2 GATATTGACGGAGTCTTCTGGGTCGCTAGT 421–450 This study R2 ATGCGCGTGAAGTAGATCTGGAATTAGGAG 584–612 This study F3 TCGCTAGTAACCAGGCTGATGTCAATACCC 442–472 This study R3 CAGAATTGGCTCTACTACGCGATCCTGCAC 629–658 This study RT-RAA-LFD Nfo probe (FAM)ACCCAAGTAGCGATGAGGCTATTCCGACTA[THF]GTTTCCGCCTGGCACGG-C3Spacer 494–541 This study F TCGCTAGTAACCAGGCTGATGTCAATACCC 442–472 This study R (Biotin) GAGCAGACCTTCCTGAGCCTTCAATATAGT 557–586 This study RT-PCR P1 GAGCGTCCTTTGGAAATCGT — [32 ] P2 GCTTAGTTACTTGCTGTGGC — [32 ] RT-qPCR P3 TCGTTCTGGTAATGGCATCCT — [33 ] P4 AGTAGCAGTTTGCTTGGGTTGAG — [33 ]
FAM-dT: thymidine nucleotide carrying fluorescein; THF: tetrahydrofuran spacer; BHQ1-dT: thymidine nucleotide carrying black-hole Quencher-1; C3 spacer: polymerase extension blocking group; FAM: carboxyfluorescein.
