Table 3.
Primer combinations and assay sensitivities assessed during the development of the Batrachovirus PCR protocols.
| Primers set | Viral species | |||
|---|---|---|---|---|
| RaHV-1 | RaHV-2 | RaHV-3 | BfHV-1 | |
| EUPB-FW-1/EUPB-RV-1 | c > 108*§ | c ≥ 106 | c > 101 (40 copies) | c > 102 (300 copies) |
| EUPB-FW-1/EUPB-RV-2 | c ≥ 106 | c ≥ 108* | 105 ≥ c > 101† | 105 ≥ c > 102 |
| EUPB-FW-1/EUPB-RV-3 | c ≥ 106 | c ≥ 108* | 105 ≥ c > 101† | 105 ≥ c > 102 |
| EUPB-FW-2/EUPB-RV-1 | c > 106§ | c ≥ 108 | c ≥ 101‡ | c ≥ 102‡ |
| EUPB-FW-2/EUPB-RV-2 | 106 ≥ c > 104† | c > 108*§ | 105 ≥ c > 101† | 105 ≥ c > 102† |
| EUPB-FW-2/EUPB-RV-3 | 106 ≥ c > 104† | c ≥ 108* | 105 ≥ c > 101† | 105 ≥ c > 102† |
| EUPB-FW-3/EUPB-RV-1 | c > 108*§ | c > 108*§ | 105 ≥ c > 101† | 105 ≥ c > 102† |
| EUPB-FW-3/EUPB-RV-2 | c ≥ 104 | c ≥ 107 | c ≥ 102 | c ≥ 103 |
| EUPB-FW-3/EUPB-RV-3 | c ≥ 105 | c ≥ 106 | c ≥ 104 | c ≥ 105 |
| EUPB-FW-4/EUPB-RV-4 | c ≥ 103‡ | c ≥ 104‡ | c ≥ 103‡ | c ≥ 103‡ |
c = copies.
*
The 108 suspensions contained 2–4 × 108 copies per microliter.
†
Values between 2 different dilutions refer to the lowest and highest dilution, which yielded a positive and negative signal, respectively. Specifically, for these primer combinations, no intermediate dilutions were tested.
‡
Recommended primer combinations along with their sensitivities.
§
No amplification was obtained in the PCR reaction using the plasmid copy number shown in the table, and no additional dilution was considered.