Application of the antigen-antibody（P24 polyclonal and monoclonal antibody） binding in detecting BDV-P24 protein in paraffin tissue sections

Abstract:

Objective：To verify the antigen-antibody（P24 polyclonal and monoclonal antibody） binding in detecting Borna disease virus（BDV）-P24 protein in paraffin tissue sections and to explore the work conditions. Methods：Forty Sprague-Dawley rats born within 24 h were intracranially（i.c.） inoculated in the right cerebral hemisphere with either BDV solution or phosphate-buffered saline（con－trol：sham inoculated）. Forty Sprague-Dawley rats and 40 C57 rats without injection were chosen as negative controls. The brains of rats on 60 d after birth were perfused and were subsequently embedded into paraffin. Then the immunohistochemical staining was per－formed following Envision two-step protocol. The primary antibody was chosen as P24 polyclonal and monoclonal antibody and the di－lution ranged from 1∶50 to 1∶5 000. The primary antibody was replaced by PBS as blank control. The brownish yellow or brown parti－cles distributing in the cells were considered as positive and total score was got by multiplying positive cell proportion score and posi－tive intensity score. Results：There were significant differences in staining scores between experimental group and control group（polyclonal P24 group Z=-3.108，P=0.000，monoclonal P24 group Z=-4.605，P=0.000. It showed a high sensitivity with slightly stained background in dilution 1∶200，1∶400 using polyclonal P24 staining while a high sensitivity without stained background in dilution 1∶100，1∶200，1∶400 using monoclonal P24 staining. Conclusion：BDV P24 polyclonal and monoclonal antibody can bind the antigen distributed in paraffin sections of BDV infectious rat. The recommended dilution of polyclonal P24 antibody is 1∶200 and 1∶400. The recommended dilution of monoclonal P24 antibody is 1∶100，1∶200 and 1∶400.