Table 2 Confirmation of variants. The confirmation of deletions was performed by both real-time PCR and conventional PCR with Sanger sequencing of PCR products. The verification of indel and SNP, both causing premature stop codon, used conventional PCR coupled with Sanger sequencing of PCR products. 18S real-time PCR [23] was performed to control for the presence/amount of H. meleagridis DNA
strain | passage number | g6116vir conv. PCR | g6116vir real-time PCR | 18S real-time PCRa | g7085vir (conv PCR, real-time PCR) | indel (AGC Kinase) | SNP (LRR) |
|---|---|---|---|---|---|---|---|
monoxenic 2922-C6/04/DH5 α | 28p | + | + (Ct 23.68) | + (Ct16.29) | + | wtc | wt |
290p | – | - (no Ct) | + (Ct18.70) | + | premature stop | premature stop | |
xenic 2922-C6/04 | 13p | + | + (27.59) | + (Ct19.87) | + | wt | wt |
51p | + | + (Ct29.93) | + (Ct17.10) | + | wt | wt | |
83p | + | + (Ct 32.25) | + (Ct21.49) | + | wt | wt | |
145p | – | - (no Ct) | + (Ct19.47) | + | premature stop | wt | |
237p | – | - (no Ct) | + (Ct18.05) | + | premature stop | premature stop | |
292p | – | - (no Ct) | + (Ct16.23) | + | premature stop | premature stop | |
xenic 2877-C3/05 | 22p | + | + (Ct25.57) | + (Ct18.31) | n.d.b | wt | wt |
xenic 8175-C7/06 | 21p | + | + (Ct26.02) | + (Ct18.07) | n.d. | wt | wt |
xenic 13,250-C20/10 | 24p | + | + (Ct24.83) | + (Ct17.27) | n.d. | wt | wt |
316p | + | + (Ct22.91) | + (Ct16.13) | n.d. | wt | wt |