Rapid Detection of Bovine Viral Diarrhea Virus in Viremic Cattle by Flow-Cytometry
DOI:
https://doi.org/10.21423/aabppro19926439Keywords:
leukocyte analysis, viremic cattle, Flow cytometry, immunofluorescence, BVDVAbstract
A fast and simple assay with high sensitivity to BVDV in blood samples independent of blood storage and clinical status of animals. is described. A pan-pesti virus reactive monoclonal antibody Cl6 (BVDV-p 80/125 specific) was applied for immunofluorescence (IF) in flow cytometry with leukocytes and for IF with infected bovine kidney cells as a reference method. From 304 blood samples of patients of the veterinary clinic 63 NCP- and 7 CP- BVDV strains were isolated. There is a high BVDVspecific IF in granulocytes and monocytes, whereas in lymphocytes the IF intensity per cell is weaker. The percentage of antigen positve cells varies from 1% to 87 % (mean 22%) in granulocytes/monocytes and 0% to 37% (mean 8%) in lymphocytes. There is a total correlation between conventional BVDV-diagnosis with tissue culture isolation and leukocyte analysis with flow cytometry. After blood freezing or storage at +8°C for 6 weeks BVDV detection in leukocytes is not significantly influenced.
Flow cytometry provides a quantitative, objective analysis of leukocyte associated BVDV-antigen. Both low levels of antigen per cell and low percentages of antigen positive cells are detectable. Direct leukocyte analysis with flow cytometry is a rapid (2 hours). easy and reliable method for detection of viremic cattle.