Detection improvement of cytomegalovirus antigen in human peripheral blood using monoclonal antibodies and automated reading of cell preparations.
One of the major drawbacks in cytomegalovirus (CMV)-antigenaemia detection for diagnosis of active CMV infection is the low number of CMV-antigen positive cells present in peripheral blood. It is therefore necessary to screen large numbers of peripheral blood granulocytes to find only a few antigen-positive cells. We have optimized this detection by testing several monoclonal antibodies (mAb) to CMV-antigens (mAbs C10/C11, C12, BM222, E13 and SL20). In total 550 blood samples from 40 patients were investigated. More blood samples were found positive with mAb C12 than with the other mAbs. Also the average number of positive cells per slide was highest for mAb C12. Furthermore, duplicate slides were examined automatically using an image analysis system (LEYTAS) and compared to visual detection (cytospin slides). The detection sensitivity of both screening methods was compared for mAb C12. In total 360 slides were analysed, from positive as well as negative blood samples. The sensitivity of the automated screening was 93% and for the visual evaluation of the cytospin slides 73%. In conclusion, mAb C12 was the most suitable of the mAbs tested for detection of antigenaemia, and automatic detection of CMV antigenaemia with image analysis of slides is a sensitive method due to the large numbers of cells that can be screened.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000956:Antigens, Viral / Q000097:blood*; D003587:Cytomegalovirus / Q000276:immunology*; D003586:Cytomegalovirus Infections / Q000175:diagnosis; D006801:Humans; D007165:Immunosuppression Therapy
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral