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Intralesional bovine papillomavirus DNA loads reflect severity of equine sarcoid disease
Authors:R. HARALAMBUS  J. BURGSTALLER  J. KLUKOWSKA‐RÖTZLER  R. STEINBORN  S. BUCHINGER  V. GERBER  S. BRANDT
Affiliation:1. Equine Clinic, VetSuisse Faculty, University of Berne, Laenggass‐Strasse 120, CH‐3012 Berne, Switzerland;2. Department of Biotechnology in Animal Production, IFA‐Tulln, Konrad‐Lorenz‐Strasse 20, A‐3430 Tulln, Austria;3. Vetomics Core Facility, University of Veterinary Medicine, Veterinaerplatz 1, A‐1210;4. Faculty of Computer Science, University of Vienna, Lenaugasse 2/8, A‐1080;5. and;6. Equine Biotechnology Unit, University of Veterinary Medicine, Veterinaerplatz 1, A‐1210 Vienna, Austria.;7. Author to whom correspondence should be addressed.
Abstract:Reasons for performing study: Sarcoids are nonmetastasising, yet locally aggressive skin tumours that constitute the most frequent neoplasm in equids. Infection by bovine papillomaviruses types 1 and 2 (BPV‐1, BPV‐2) has been recognised as major causative factor in sarcoid pathogenesis, but a possible correlation of intralesional virus load with disease severity has not been established thus far. Hypothesis: Given the pathogenic role of BPV‐1 and BPV‐2 in sarcoid disease, we suggest that intralesional viral DNA concentration may reflect the degree of affection. Methods: Severity of disease was addressed by recording the tumour growth kinetics, lesion number and tumour type for 37 sarcoid‐bearing horses and one donkey. Viral load was estimated via quantitative real‐time PCR (qPCR) of the E2, E5, L1 and L2 genes from the BPV‐1/‐2 genome for one randomly selected lesion per horse and correlated with disease severity. Results: Quantitative PCR against E2 identified viral DNA concentrations ranging from 0–556 copies/tumour cell. Of 16 horses affected by quiescent, slowly growing single tumours or multiple mild‐type lesions, 15 showed a viral load up to 1.4 copies per cell. In stark contrast, all equids (22/22) bearing rapidly growing and/or multiple aggressive sarcoids had a viral load between 3 and 569 copies per cell. Consistent results were obtained with qPCR against E5, L1 and L2. Conclusions: While tumours of the same clinical type carried variable virus load, confirming that viral titre does not determine clinical appearance, we identified a highly significant correlation between intralesional viral load and disease severity. Potential relevance: The rapid determination of BPV viral load will give a reliable marker for disease severity and may also be considered when establishing a therapeutic strategy.
Keywords:horse  bovine papillomavirus  equine sarcoid  quantitative PCR  viral DNA load
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