149-P: TIME AIN’T ON OUR SIDE: REAL-TIME PCR FOR DECEASED DONOR TYPINGS

Aim A string of problem typings of three (3) deceased donors led us to conclude that the tried-and-true method of SSP may no longer be optimal for such typings. The necessity of running parallel SSP kits and the well-known problems with safety and disposal of EtBr-treated gels also led us to consider real-time PCR for deceased donor HLA typing. Methods We decided to compare side by side our current SSP method for deceased donor typing with real time PCR HLA typing utilizing Linkage Biosciences LinkSēq kits with Sure Typer analytic software. Results An average SSP typing for a deceased donor takes about 4 hours and the LinkSēq kits takes approximately 2 hours. SSP typing required the running of an additional kit from another vendor to provide timely backup in the event of a mistyping or drop out. There have been, to date, no need to repeat testing or have additional backup kits run with the real-time PCR HLA typing. Unfortunately, while the Linkage Biosciences kits are competitively priced with SSP, the necessity of buying a real-time PCR instrument may be prohibitive for many laboratories. Conclusions We had decided to run a second SSP tray for both Class I and II antigens in order to prevent donor mis-typings. This was also done in order to prevent the need to repeat a typing due to a failed primer/well thereby potentially adding more ischemia time to organs. However, with Real-time PCR there was a reduction in time to an average of 2 hours. When you factor in that tech time is the most expensive component of any clinical test, an appreciable reduction in expenses results. Real-time PCR gave more information to the benefit of all the patients involved. With half the time, and no additional Taq polymerase, or DNA we were able, with Real-time PCR, to type for HLA-A, -B, - C, -DRB1, -DRB3, 4, 5, and -DQB1 antigenic equivalents, and, in addition, -DQA1. In conclusion, we could not ignore the time savings, the cost savings, and just overall how much better it was for our patients.