Veterinary Archives, Vol. 91 No. 5, 2021.
Original scientific paper
https://doi.org/10.24099/vet.arhiv.1047
Establishment of a sandwich ELISA for bovine plasma PON-1 and its predictive capabilities for dairy cows with fatty liver
Chang Zhao
; College of Animal Science and Technology, Anhui Agricultural University, Hefei, PR China
Yunlong Bai
; College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, PR China
Dong Wang
; Heilongjiang Provincial Key Laboratory of Prevention and Control of Bovine Diseases, College of Animal
Cheng Xia
; College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, PR China
Abstract
The aim of this study to improve the clinical diagnosis of fatty livers (FL) in dairy cows by using the paraoxonase-1 (PON-1) enzyme as a detection index. Prokaryotic expression technology was used to generate recombinant bovine PON-1 protein. Mice were immunized with this protein to generate hybridoma cells, stably secreting anti-PON-1. Cells were injected into the peritoneal cavity of mice, and ascites were purified to generate bovine PON-1 monoclonal antibody. Rabbits were then immunized with this antigen, and a polyclonal antibody against bovine PON-1 was obtained. Using monoclonal and polyclonal antibodies, a double-antibody sandwich ELISA for plasma PON-1 was constructed. Plasma samples were collected from healthy (n = 13) and FL (n = 13) cows, and plasma PON-1 levels were detected using the PON-1 ELISA. Receiver operating characteristic curve (ROC) analysis was used to analyze correlations between PON-1 levels and FL. Results showed that the ideal working concentration of the monoclonal antibody was 0.8 mg/mL, and the quantitative detection limit was 90 ng/mL. Plasma PON-1 levels were significantly lower in FL cows, when compared with healthy animals. It is concluded that PON-1 ELISA predicts risk factors for dairy cows with FL. PON-1 levels in plasma can be used as an early warning indicator for FL and concentration of 61.87 nmol/L was identified as warning index.
Keywords
clinical applications; ELISA; fatty liver; PON-1; ROC
Hrčak ID:
271609
URI
Publication date:
12.12.2021.
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