1012-LB Enhanced resolution of LIFECODES HLA-C eRES SSO typing kit by the addition of 22 new probes

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LIFECODES SSO typing kits are based upon the Luminex platform to provide intermediate level of resolution mostly focused upon resolving common well documented (CWD) HLA alleles. Out of 81 CWD HLA-C alleles identified by Cano et al (Human Immonol. 68: 392-417, 2007), 71 CWD alleles are unique at the 4-digit level. An analysis of the CWD-CWD allelic resolution by LIFECODES HLA-C product revealed that 14 CWD-CWD alleles could not be resolved. The aim of the study was to develop probes to resolve these and other ambiguities. Herein, we demonstrate enhanced resolution of the kit by the addition of 22 new probes. Sequence specific probes, directed to nucleotide polymorphisms in exons 2 and 3 of HLA-C genes were designed to resolve various ambiguities of the CWD alleles. These probes were added to current HLA-C probe mix to evaluate the enhanced resolution of the revised kit. Addition of 22 new probes to the LIFECODES HLA-C probe mix resulted in the incremental resolution of 10 CWD-CWD, 12 CWD-rare and 2 rare-rare allelic ambiguities. For CWD-CWD resolution, Probe-276 resolved C*04:01 from C*04:07; Probe-280 resolved C*03:10 from C*03:09; Probe-373 resolved C*03:05 from C*03:04; Probe-376 resolved C*05:05 from C*05:01; Probe-378 resolved C*15:09 from C*15:04; Probe-380 resolved C*03:06 from C*0302; Probe-382 resolved C*12:13 from C*12:03; Probe-383 resolved C*07:12 from C*07:04; Probe-384 resolved C*01:08 from C*01:02; and Probe-385 resolved C*08:06 from C*08:03. For allelic combinations, Probe-276 resolved C*04:01 +C*07:02 pair from C*07:64 + C*18:01/02 pair; Probe 279 resolved C*05:20/32 + C*07:01 pair from C*07:01 + C*08:02 pair; Probe-381 resolved C*12:03 + C*07:01 pair from C*06:11 + C*07:92 pair, and C*12:03 + C*07:40 pair from C*07:01 + C*16:04 pair. Other probes resolved C*01:02/01:07, C*01:02/01:18, C*02:04/02:02, C*02:10/02:14, C*04:01/04:05, C*06:02/06:15, C*07:05/07:27, C*07:14/07:27, C*07:35/07:55N, and C*17:04/17:01 allelic ambiguities. Out of 178 samples typed, 123 (69%) were resolved to a single pair of CWD alleles at 2-field resolution over exons 2 and 3. Efforts are underway to resolve the remaining CWD ambiguities.