Establishment of a Visual Gene Chip Method for the Simultaneous Detection of Seven Waterfowl Virus Pathogens

Figures

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  Figure 1. Arrangement of chips. Biotin is the positive QC for the gene chip; buffer is the negative QC for the gene chip.

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  Figure 2. PCR products of recombinant positive plasmids by agarose gel electrophoresis. M: marker; 1: GPVXP; 2: DEVXP; 3: MDPVXP; 4: DHAV-3XP; 5: DTMUVXP; 6: DHAV-1XP; 7: NDRVXP; NC: negative control.

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  Figure 3. Optimisation of the annealing temperature for the PCR amplification. M: marker; 1: 54 °C; 2: 56 °C; 3: 58 °C; 4: 60 °C; 5: 54/56 °C; 6: 54/58 °C; 7: 56/58 °C; 8: 56/60 °C; NC: negative control.

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  Figure 4. Optimisation of PCR amplification extension times. M: marker; 1: 10 s; 2: 15 s; 3: 20 s; 4: 30 s; NC: negative control.

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  Figure 5. Optimisation of primer concentrations for PCR amplification. M: marker; 1: 0.05 μmol/L; 2: 0.1 μmol/L; 3: 0.2 μmol/L; 4: 0.4 μmol/L; NC: negative control.

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  Figure 6. Optimisation of hybridisation times for gene chips.

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  Figure 7. Optimisation of SA-HRP concentration for gene chips.

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  Figure 8. Specificity of the gene chip. (A): GPV; (B): DEV; (C): MDPV; (D): DHAV-1; (E): DHAV-3; (F): DTMUV; (G): NDRV; (H): Dastv; (I): DadV-3; (J): negative control.

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  Figure 9. Sensitivity of single plasmid standards for gene chips. (A): 1.0 × 103 copies/μL; (B): 1.0 × 102 copies/μL; (C): 1.0 × 101 copies/μL; (D): 1.0 × 100 copies/μL; (E): negative control.

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  Figure 10. Sensitivity of mixed plasmid standards for gene chips. (A): 1.0 × 104 copies/μL; (B): 1.0 × 103 copies/μL; (C): 1.0 × 102 copies/μL; (D): 1.0 × 101 copies/μL; (E): 1.0 × 100 copies/μL.

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  Figure 11. Stability of the gene chip. (A): 0 days; (B): 30 days; (C): 60 days; (D): 90 days; (E): 120 days; (F): 180 days.