COMPARISON OF HIGHER RESOLUTION OF SEQUENCE SPESIFIC OLIGONUCLEOTIDE PROBE vs NEXT GENERATION SEQUENCING FOR HLA A, B, C AND DRB1 TYPİNG
Project no. 31968 · Status: Kapanmış (Yayın bekleniyor) · 2023
Summary
<p>Human Leukocyte Antigen (HLA) compatibility is crucial for the long-term success of kidney transplants. The necessity of performing high-resolution HLA typing has been recognized with the importance of eplet analysis in kidney transplants. The objective of the study is to determine the accuracy and the level of resolution of higher resolution of sequence spesific oligonucleotide probe (SSO) vs next generation sequencing (NGS). Twenty-eight samples from healthy bone marrow donors and 10 HR BEPT 2024 samples were included in the study. DNA samples were extracted using Maxwell RSC Whole Blood DNA Kit (Promega). HLA typing was done using LabType XR Class I A, B, C and Class II DRB1 Typing Test (One Lambda) and AllType FASTPlex NGS 11 Loci Flex Kit (OneLambda). All tests were done according to manufacturer’s instructions using LabScan3D for SSO and iseq 100 (illumina) for NGS. Our study was funded by Ege University Office of Scientific Research Projects with project number 31968. A total of 304 alleles from 38 samples were typed with both methods for HLA-A,B,C and DR loci. The results were concordant with NGS at low resolution except for two (0.65%) assingments for one specimen, which produced no result for HLA-C. High resolution typing results were concordant except for two (0,65%) ambiguities and two (0.65%) assignments among three samples. The HLA-A*23:01 NGS result was A*23:CJT(01/17) with SSO and HLA DRB1*14:54 with NGS was DRB1*14:BCAD (01/54). Low resolution HLA-B result could be obtained for one sample (B*27 B*35). All discordant results were tested in duplicate. Among 304 alleles, 6 (1.97%) either produced no result or was discordant with NGS.NGS should be used where available for highly sensitized kidney transplant candidates. Higher resolution SSO may be helpful when NGS isn’t available and for deceased donors. </p>