Development of Rapid Isothermal Detection Methods Real-Time Fluorescence and Lateral Flow Reverse Transcription Recombinase-Aided Amplification Assay for Bovine Coronavirus

Figure 6

BCoV real-time RT-RAA sensitivity, specificity, and reproducibility assays. (a) Sensitivity assay. The prepared standard RNA was diluted to concentrations ranging from 1.46 × 10⁷ to 1.46 × 10⁰ copies/μL. Water without DNAzyme served as the negative control. Real-time RT-RAA was able to detect as low as 1.46 × 10¹ copies/μL. (b) Specificity assay. Nucleic acids from BCoV, BVDV, BEV, BPIV3, BEFV, and BRV were used as templates. Water without DNAzyme was used as a negative control. Real-time RT-RAA demonstrated specific amplification only for BCoV. (c) Reproducibility Assay. Reproducibility experiments were conducted using three standard plasmid dilutions: 1.46 × 10⁷, 1.46 × 10⁴, and 1.46 × 10³ copies/μL.

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