Bovine Coronavirus (BCoV) is a significant pathogen responsible for neonatal calf
diarrhea, winter dysentery in adult cattle, and bovine respiratory diseases. Infection
with the virus can result in hemorrhagic diarrhea, decreased milk production, and
potentially fatal outcomes in cattle, leading to considerable economic repercussions
for the cattle industry. Efficient management of BCoV relies on swift and precise
detection techniques. CHO cells were utilized to express a secreted recombinant
nucleocapsid protein (N), whereby rabbit polyclonal antibodies (pAb) were generated
through immunization. An indirect enzyme-linked immunosorbent assay (iELISA)
based on N protein was established for the de... More
Bovine Coronavirus (BCoV) is a significant pathogen responsible for neonatal calf
diarrhea, winter dysentery in adult cattle, and bovine respiratory diseases. Infection
with the virus can result in hemorrhagic diarrhea, decreased milk production, and
potentially fatal outcomes in cattle, leading to considerable economic repercussions
for the cattle industry. Efficient management of BCoV relies on swift and precise
detection techniques. CHO cells were utilized to express a secreted recombinant
nucleocapsid protein (N), whereby rabbit polyclonal antibodies (pAb) were generated
through immunization. An indirect enzyme-linked immunosorbent assay (iELISA)
based on N protein was established for the detection of BCoV antibodies. Reaction
conditions were optimized using a checkerboard approach, with the optimal
antigen concentration at 1.25 μg/mL and the optimal antibody dilution at 1:200,
the cutoff value distinguishing negative and positive serum samples was 0.986. The
sensitivity test indicated that this rabbit pAb had a maximum dilution of 218 within
the assay range, did not cross-react with BHV-1, BVDV, BRV, and BRSV positive
serum samples, and shown great specificity. The developed iELISA method and
commercial kit were used to test 58 bovine serum samples, and the concordance
rate was 94.83%. In summary, we have developed a cost-efficient and precise
iELISA method based on N protein that serves as a useful diagnostic tool for BCoV
in clinical samples and epidemiological research.
