BackgroundBovine leukemia virus (BLV) is associated with enzootic bovine leukosis, which is the most common neoplastic disease of cattle. BLV infects cattle worldwide, imposing a severe economic impact on the dairy cattle industry. Recently, we developed a new quantitative real-time polymerase chain reaction (PCR) method using Coordination of Common Motifs (CoCoMo) primers to measure the proviral load of known and novel BLV variants in BLV-infected animals. Indeed, the assay was highly effective in detecting BLV in cattle from a range of international locations. This assay enabled us to demonstrate that proviral load correlates not only with BLV infection capacity as assessed by syncytium formation, but also with BLV disease progression. In this study, we compared the sensitivity of our BLV-CoCoMo-qPCR method for detecting BLV proviruses with the sensitivities of two real-time PCR systems, and also determined the differences of proviral load with serotests.ResultsBLV-CoCoMo-qPCR was found to be highly sensitive when compared with the real-time PCR-based TaqMan MGB assay developed by Lew et al. and the commercial TaKaRa cycleave PCR system. The BLV copy number determined by BLV-CoCoMo-qPCR was only partially correlated with the positive rate for anti-BLV antibody as determined by the enzyme-linked immunosorbent assay, passive hemagglutination reaction, or agar gel immunodiffusion. This result indicates that, although serotests are widely used for the diagnosis of BLV infection, it is difficult to detect BLV infection with confidence by using serological tests alone. Two cattle were experimentally infected with BLV. The kinetics of the provirus did not precisely correlate with the change in anti-BLV antibody production. Moreover, both reactions were different in cattle that carried different bovine leukocyte antigen (BoLA)-DRB3 genotypes.ConclusionsOur results suggest that the quantitative measurement of proviral load by BLV-CoCoMo-qPCR is useful tool for evaluating the progression of BLV-induced disease. BLV-CoCoMo-qPCR allows us to monitor the spread of BLV infection in different viewpoint compared with classical serotest.
SUMMARYTwo-day-old specific-pathogen-free chicks were inoculated intranasally with the MA-87 strain of infectious bronchitis virus, and trachea and kidney lesions studied histologically and immunohistochemically. Lesions and viral antigen were first detected in the trachea; severe damage was then observed in the kidney. Viral antigen appeared prior to the development of renal lesions and was detected in the cytoplasm of epithelial cells by 4 days post-inoculation (p.i.). The epithelial cells of the collecting ducts, collecting tubules and distal convoluted tubules were first affected, followed by involvement of Henle's loops, whereas the proximal convoluted tubules were only minimally affected. Antigen-positive cells of ducts and tubules were degenerated and desquamated. The severe epithelial cell damage resulted in infiltration of heterophils and macrophages in the interstitium, ducts and tubules. The detection of viral antigen was consistent with the distribution of histological lesions at 6 to 8 days p.i. At a later stage, antigen-positive cells disappeared and repair of epithelial cells was seen, accompanied by interstitial lymphoplasmacytic infiltration and lymphoid nodular formation.
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