Establishment of a reverse transcription recombinase-aided amplification detection method for porcine group a rotavirus

Figures

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  Figure 1. Schematic diagram of primer amplification. (A) Schematic diagram of the RT-RAA primer screening. The numbers in the primer name represent the position of the oligonucleotides in the conservative region of the VP6 gene. (B) Schematic diagram of Taqman probe-based RT-qPCR primers amplification. The numbers in the primer name represent the position of the oligonucleotides in the conservative region of the VP6 gene.

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  Figure 2. (A) Best primer screen. M: DL2000 marker; 1–9: 9 pairs of PoRVA primers, 1: R-F1-R1, 2: R-F1-R2, 3: R-F1-R3, 4: R-F2-R1, 5: R-F2-R2, 6: R-F2-R3, 7: R-F3-R1, 8: R-F3-R2, 9: R-F3-R3; 10: Negative Control. (B) RT-RAA primer screening results. :R-F1-R1, : R-F1-R2, : R-F1-R3, : R-F2-R1, : R-F2-R2, : R-F2-R3, : R-F3-R1, : R-F3-R2, : R-F3-R3; : negative control. TT value: : 6.16, : 7.84, : 9.19, : 9.34, : 9.27, : 10.80, :17.72, : 18.96, : 20.20, : none.

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  Figure 3. (A) The results of RT-RAA sensitivity analysis: RT-RAA PoRVA plasmid as standard, -: 1 × 105-1 × 10−1 copies per reaction sequentially; : negative control. (B) The results of TaqMan probe-based RT-qPCR sensitivity analysis: TaqMan probe-based RT-qPCR PoRVA plasmid as standard, -: 1 × 107-1 × 1 0−1 copies per reaction sequentially; : negative control.

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  Figure 4. Specificity of the RT-RAA assay for PoRVA. -: PoRVA, PRRSV, JEV, APPV, SVV, CSFV, and GETV virus RNA, respectively; : negative control.