Skin tissue samples from all groups following different sites of inoculation with ORFV isolates showed positive PCR amplification DNA bands at 1062 bp (representing F1L gene) and 1199 bp (representing B2L gene) in size (Figure 8).
In this study, ORFV was detected from the skin samples of experimentally infected mice through PCR amplification targeting at two important genes which were B2L and F1L.
(Lanes 1, 3, 5, 7, 9,11,13, and 15: F1L primer; Lanes 2,4,6,8,10,12,14, and 16: B2L primer).
We assessed the performance of the PPV real-time PCR assay with, as calibrators, 4 plasmids that were cloned according to routine procedures and contained 487 by of the B2L gene of ORFV (strain B006, pORFV), PCPV (strain B021, pPCPV), BPSV (strain B177, pBPSV), and SPV (pSPV), including the original sequence of these isolates in the primer-binding (PPV up, PPV do) and probebinding (PPV TMGB) region (cloning primers are shown in supplemental material Table 1 in the online Data Supplement).
Although there is a divergence of up to 15.6% in the B2L gene sequences of different PPV species, careful selection of primers and probe gave satisfactory and comparable amplification efficiency as well as a detection limit for the PPV species ORFV, BPSV, PCPV, and SPV, which was confirmed by analysis of 41 clinical samples from humans or animals infected with these PPV species.
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PCR was used to amplify the PPV B2L gene encoding a major envelop protein (3,5), and a specific amplification product has been obtained from all the isolates.
Surprisingly, the B2L sequences of the 3 red deer isolates showed 100% identity with that of PVNZ RD86, suggesting that PVNZ could be the cause of the disease.
The research was based on answers from 3,079 landlords who responded to a questionnaire in B2L
Model BIL and B2L
Budget hopper loaders have an easy-to-use reset circuit breaker on outside of control box.