161-P: REAL TIME (AND COST) SAVINGS WITH REAL TIME PCR DECEASED DONOR MOLECULAR TYPING

Aim UNOS requires that deceased donor (DD) HLA typing be performed by molecular methods. Antibodies to more and varied HLA alleles are detected by Luminex single antigen beads making crossmatch interpretation potentially difficult. The aim of this study was to evaluate the time and cost of performing DD molecular typing with Real Time PCR. Methods We compared the cost and time it takes to obtain a low resolution DNA typing by our current method standard PCR-SSP/Serology typing to the Real–Time (RT) PCR typing by Linkage Biosystems. Eight previously DNA typed samples were tested by RT PCR kit HLA-ABCDRDQA1DQB1DP+ kit. RT PCR uses the melt curve analysis to identify different alleles. Our current deceased donor typing method includes class I and II testing for HLA A,B,C,DRB1,DRB3,4,5 and DQB using serologic and molecular techniques. We use both methods to guard against no-amps by PCR and to identify any nulls alleles. Typing kits, reagents and tech time were included in the total cost analysis. Results All the HLA typings agreed. DNA isolation was required for both assays and was not included in the comparison [ Table 1 ]. Tech time savings with RT PCR was 140 minutes. The increased tech time for Std PCR typing was due to the need for multiple kits. The RT PCR software analysis streamlines the antigen assignment by identifying rare alleles. The cost to run the null trays and test for DP, and DQA alleles using Std PCR which are all available on one RT PCR kit, was $419 and doubled the tech time. Conclusions RT PCR offers a time and cost efficient way to perform low resolution molecular DD typing. In our lab, Std PCR is still necessary for higher resolution typing.