Abstract:

The early diagnosis of bovine pregnancy is very important. Pregnancy-associated glycoprotein 6 (PAG6) is a specific protein secreted by maternal placental tissue to the peripheral circulation. Therefore, this study is to prepare a test strip for rapid detection of bovine pregnancy-associated glycoprotein 6. The polyclonal antibody against bovine pregnancy-associated glycoprotein 6 was prepared and diluted and added to the NC membrane. A rapid detection strip for boPAG6 was prepared by fixing and sealing. The serum to be tested was added to the test strip for incubation and then washed, and then the second antibody was added for washing. Finally, the chromogenic solution was added for rapid detection. The standardized rapid test strip diluted the antibody to 10 μg/mL with PBS and put it on the NC film. The antibody was fixed at 37℃ for 30 min and sealed with 5% skim milk powder. The serum to be tested was dropped onto the test strip and incubated at 37℃ for 1 h, then washed and secondary antibody was incubated. Finally, DAB chromogenic kit was used for 3 min to determine the detection. The preliminary application in clinical detection showed that pregnancy could still be determined in the mixed serum of non-pregnant cows and pregnant cows, and the specificity of rapid detection of the test strip was strong. The sensitivity of the rapid detection of the test strip was good at the minimum detection limit of 3200 times dilution of serum. The coincidence rate with ELISA was above 80%. The experiment has good repeatability and the test strip can be stored at 4℃ and -20℃ for 2 weeks. The test strip for rapid detection of early pregnancy diagnosis in dairy cows is established and can be applied to preliminary pregnancy diagnosis.

Key words: dairy cows, early pregnancy, diagnosis, pregnancy-associated glycoprotein 6, test strip for rapid detection