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Table II.
Applied PCR cycles for the detection of genus- and species-specific Brucella and differentiation between vaccine and field strains.
| Initial denaturation | No. of cycles | Denaturation | Annealing | Extension | Post-PCR step | |
|---|---|---|---|---|---|---|
| Genus-specific identification | 95 °C for 5 min | 40 | 94 °C for 30 s | 56 °C for 30 s | 72 °C for 1 min | 72 °C for 7 min |
| Differentiation of Brucella species | 95 °C for 35 s | 45 | 95 °C for 30 s | 58 °C for 30 s | 72 °C for 45 s | 72 °C for 10 min |
| Differentiation between vaccine and field strains | – | 40 | 95 °C for 1 min | 62 °C for 1 min | 72 °C for 1.5 min | – |