Abstract
Aim Proper selection of positive control (PC) serum in flow cytometry (FC) crossmatch is essential in validation the procedure. In our laboratory we developed a selection method of the optimal pooled PC serum dilution that assures the sensitivity superiority of flow crossmatch over anti-human globulin enhanced complement dependent cytotoxicity (CDC-AHG). Methods Highly and broadly sensitized patients with >10000 MFI were pooled together. The pooled PC is serially diluted (1:2-1:256) and CDC-AHG crossmatched against target cells in which the end point was determined (highest dilution giving positive reaction). Thereafter, we FC crossmatched target cells with the further diluted PC serum than end point observed in the CDC-AHG crossmatch. Results Positive reactions were seen at 1:8 dilutions in CDC-AHG crossmatch, however; they were positive up to 1:128 in FC crossmatch. Therefore, we selected the highest dilution of pooled PC serum that shows positive flow cytometry crossmatch. Conclusions In flow cytometry crossmatch, using optimized dilution of the positive control rather than extremely positive control serum, will assure the flow cytometry crossmatch sensitivity superiority over CDC-AHG crossmatch.
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