HLA-specific qPCR assays developed in this study
| Target HLA locus . | Assay name . | Target HLA allele groups† . | Efficiency (%)†,‡ . | Sensitivity (%)†,§ . | Informative HSCT (%)‖ . | |
|---|---|---|---|---|---|---|
| Haploidentical . | Unrelated . | |||||
| (n = 454) . | (n = 113) . | |||||
| A* | KMR501-A | 01, 36 | 87.7 | 0.03 | 10.4 | 0 |
| KMR502-A | 02 | 87.6 | 0.03 | 12.1 | 0 | |
| KMR504-A | 11 | 94.4 | 0.16 | 3.5 | 0.8 | |
| KMR505-A | 23, 24 | 98.0 | 0.03 | 9.9 | 0.8 | |
| KMR506-A | 25, 26, 34, 66, 68, 69 | 94.3 | 0.16 | 11.2 | 4.4 | |
| C* | KMR511-C | 03 | 90.8 | 0.03 | 3.1 | 0 |
| KMR512-C | 04 | 84.5 | 0.16 | 10.6 | 0 | |
| DPB1* | KMR520-DPB1 | 04:01, 23:01, 31:01, 34:01 | 98.3 | 0.03 | 14.1 | 23.8 |
| KMR521-DPB1 | 02:01, 04:02, 08:01, 105:01, 16:01 | 100.8 | 0.03 | 17.2 | 31.8 | |
| KMR522-DPB1 | 01:01, 11:01, 13:01, 15:01 | 96.8 | 0.03 | 3.1 | 16.8 | |
| Target HLA locus . | Assay name . | Target HLA allele groups† . | Efficiency (%)†,‡ . | Sensitivity (%)†,§ . | Informative HSCT (%)‖ . | |
|---|---|---|---|---|---|---|
| Haploidentical . | Unrelated . | |||||
| (n = 454) . | (n = 113) . | |||||
| A* | KMR501-A | 01, 36 | 87.7 | 0.03 | 10.4 | 0 |
| KMR502-A | 02 | 87.6 | 0.03 | 12.1 | 0 | |
| KMR504-A | 11 | 94.4 | 0.16 | 3.5 | 0.8 | |
| KMR505-A | 23, 24 | 98.0 | 0.03 | 9.9 | 0.8 | |
| KMR506-A | 25, 26, 34, 66, 68, 69 | 94.3 | 0.16 | 11.2 | 4.4 | |
| C* | KMR511-C | 03 | 90.8 | 0.03 | 3.1 | 0 |
| KMR512-C | 04 | 84.5 | 0.16 | 10.6 | 0 | |
| DPB1* | KMR520-DPB1 | 04:01, 23:01, 31:01, 34:01 | 98.3 | 0.03 | 14.1 | 23.8 |
| KMR521-DPB1 | 02:01, 04:02, 08:01, 105:01, 16:01 | 100.8 | 0.03 | 17.2 | 31.8 | |
| KMR522-DPB1 | 01:01, 11:01, 13:01, 15:01 | 96.8 | 0.03 | 3.1 | 16.8 | |
Efficiency and sensitivity were tested on HLA-typed B lymphoblastoid cell lines obtained from the European Collection of Animal Cell Cultures or from healthy HLA-typed individuals (supplemental Table 3). For assays targeting multiple HLA allele groups, the most frequent HLA allele group in worldwide populations15 (supplemental Table 1) was used as target-positive reference DNA. Efficiency and sensitivity were determined by serial dilutions of target-positive reference DNA in water.
Reported percentages are the mean of at least 3 independent experiments performed at either of the 2 testing sites (Ospedale San Raffaele [OSR] and Universitätsklinikum Essen [UKE]). Standard error of mean was <4 in all cases.
Maximal reproducible sensitivity determined in at least 3 independent experiments performed at either of the 2 testing sites (OSR and UKE).
Percentage of informative HSCT based on HLA typing of patients and donors for 454 haplo-HSCT performed between years 2000 and 2016 at OSR, and for 113 UD-HSCT performed in 2015 at UKE with at least 1 HLA-A, HLA-C, or HLA-DPB1 mismatch in the graft-versus-host direction. Some transplants had more than 1 informative reaction, whereas others had none. The total percentages of transplants with at least 1 informative reaction was 71.6% and 66.4% for haplo-HSCT and UD-HSCT, respectively.