ABSTRACT
Bovine papillomaviruses (BPV) induce benign tumours of the cutaneous or mucosal epithelia in cattle, but are also involved in the development of cancer of the urinary bladder and of the upper gastrointestinal tract. Current BPV genotyping assays employ techniques developed originally for the detection of human papillomaviruses. These methods rely on consensus PCR amplification and subsequent sequencing and are cumbersome and limited in their analytic sensitivity to detect BPV, especially in multiple infections. In this study, a novel multiplex BPV genotyping assay is described to detect sensitively and specifically BPV-1 to -10 as well as BaPV-11. The assay is based on a multiplex PCR using novel broad-spectrum bovine papillomavirus (BSBP) primers followed by multiplex bovine genotyping (MBG) by Luminex xMAP technology. The detection limit of the assay was shown to be between 10 and 100 BPV genomes. In a first application, BPV was detected in 100% of wart preparations with BPV-8 being most prevalent, followed by types 6, 1 and 10. The majority of warts were positive for at least four BPV types. In conclusion, BSBP-PCR/MBG is a powerful high-throughput method suitable for the study of the natural history of BPV and could be useful to veterinarians for the monitoring of the efficacy of future BPV vaccines.
Subject(s)
Cattle Diseases/virology , Deltapapillomavirus/genetics , Deltapapillomavirus/isolation & purification , Papillomavirus Infections/veterinary , Polymerase Chain Reaction/methods , Warts/veterinary , Animals , Cattle , DNA Primers , DNA, Viral/analysis , DNA, Viral/genetics , Deltapapillomavirus/classification , Genotype , Limit of Detection , Oligonucleotide Probes , Papillomavirus Infections/virology , Warts/genetics , Warts/virologyABSTRACT
OBJECTIVE: We sought to evaluate whether soft-copy reading of simulated pulmonary chest lesions is influenced by ambient light and automatic optimization of cathode ray tube (CRT) monitor luminance. MATERIALS AND METHODS: Four types of simulated lesions (nodules, lines, micronodules, and patchy opacities) were superimposed over an anthropomorphic chest phantom. Lesion detection with soft-copy reading was assessed using a high-contrast grayscale 2K CRT monitor under the following conditions: (1) subdued lighting (<50 lux); (2) normal lighting conditions (450 lux) without, and (3) with a sensitivity modulation to automatically adjust the CRT luminance to the increased amount of ambient light. Reading data were analyzed according to receiver operating curve. Significance of differences was tested using an analysis of variance for repeated measures. RESULTS: Ambient room light of 450 lux did not significantly influence the detection of nodules and patchy opacities. However, bright ambient light significantly decreased detection of micronodules (0.60 vs. 0.74) and lines (0.52 vs. 0.66) relative to subdued lighting conditions. Automatic luminance adjustment could compensate the effect of ambient light for the micronodules (0.77) but not for the lines (0.53). CONCLUSION: Bright ambient light significantly decreases detection of small low-contrast structures. This may be partially but not completely compensated by an automatic luminance adaptation.
