Table 1.
Primers for PCR-SSP and TaqMan™-PCR assays
| Antigen system | dbSNP No. | Primer sequences for PCR-SSP (5′−3′) | Specificity | TaqMan™ PCR assay* |
|---|---|---|---|---|
| MNS (MN) | rs7687256, | rev: GGCAAGAATTCCTCCATAGTAG | GYPA | (self-designed assay1) |
| rs7658293 | for: CATATCAGCATTAAGTACCACTGGT | c.71G, c.72T | ||
| for: CATATCAGCATTAAGTACCACTGAG | c.71A, c.72G | |||
| MNS (Ss) | rs7683365 | for: caaaatatgattaagaaaaggaaacccg | GYPB | C__34183121_10 |
| rev: cgatggacaagttgtccca | c.143T | |||
| rev: cgatggacaagttgtcccg | c.143C | |||
| P1PK (P1/P2) | rs2143918 | rev: CACAAAGAACCTGGCTTCTCG | A4GALT | C__16072990_20 |
| for: CACATCTTTCCTGGGAAGGAATT | +2857T | |||
| for: CACATCTTTCCTGGGAAGGAATG | +2857G | |||
| LU (Lua/b) | rs28399653 | for: CCAGGGAGACCCATAACAAG | BCAM | C__25764173_10 |
| rev: TCTCAGCCGAGGCTAGGT | c.230A | |||
| rev: TCTCAGCCGAGGCTAGGC | c.230G | |||
| LU (LU8/14) | rs28399656 | rev: GAGGTCAAAGGCCAGCACAG | BCAM | C__32363989_10 |
| for: CTCTCCCAGAGGGCTACAT | c.611T | |||
| for: GATCTCTCCCAGAGGGCTACAA | c.611A | |||
| LU (Aua/b) | rs1135062 | rev: ggttaagctatggtggtcattgc | BCAM | C__1846428_10 |
| for: ccatgtcttccacttcggca | c.1615A | |||
| for: ccatgtcttccacttcggcg | c.1615G | |||
| KEL (K/k) | rs8176058 | for: GGGAGATGGAGATGGAAATGG | KEL | C__1719_20 |
| rev: GACTCATCAGAAGTCTCAGCG | c.578C | |||
| rev: GACTCATCAGAAGTCTCAGCA | c.578T | |||
| KEL (Kpa/b) | rs8176059 | rev: AGGGCACTAGGAGGAAGAAG | KEL | C__25596888_20 |
| for: CTTGTCAATCTCCATCACTTCAC | c.841C | |||
| for: CTTGTCAATCTCCATCACTTCAT | c.841T | |||
| KEL (Jsa/b) | rs8176038 | rev: ggcccttgacacttgcatac | KEL | C__25596899_20 |
| for: tgcctgggggctgcct | c.1790T | |||
| for: tgcctgggggctgccc | c.1790C | |||
| KEL (11/17) | rs61729034 | rev: CCTTAGAGGAGGGACACAAAG | KEL | C__27862879_10 |
| for: GGCAAGCTCTTCCAGATGGT | c.905T | |||
| for: GGCAAGCTCTTCCAGATGGC | c.905C | |||
| FY (Fya/b) | rs12075 | rev: GGCACCACAATGCTGAAGAG | DARC | C__2493442_10 |
| for: CTTCCCAGATGGAGACTATGG | c.125G | |||
| for: CTTCCCAGATGGAGACTATGA | c.125A | |||
| FY (Fy0) | rs2814778 | rev: CAAAGGGAGGGACACAAGAG | DARC | C__15769614_10 |
| for: CCTCATTAGTCCTTGGCTCTTAT | -67T | |||
| for: CCTCATTAGTCCTTGGCTCTTAC | -67C | |||
| JK (Jka/b) | rs1058396 | rev: AGACAGCAAGTGGGCTCAAG | SLC14A1 | C__1727582_10 |
| for: TCTTTCAGCCCCATTTGAGG | c.838G | |||
| for: GTCTTTCAGCCCCATTTGAGA | c.838A | |||
| DI (Dia/b) | rs2285644 | rev: TCCTGCCTGCCCTAGTTCTG | SLC4A1 | C__26654865_10 |
| for: GGGTGGTGAAGTCCACGCT | c.2561T | |||
| for: GGTGGTGAAGTCCACGCC | c.2561C | |||
| DI (Wra/b) | rs75731670 | rev: TGGGAGAATGCCAGGGAAAG | SLC4A1 | C__2259533_10 |
| for: CACTGGGCTTGCGTTCCA | c.1972A | |||
| for: CACTGGGCTTGCGTTCCG | c.1972G | |||
| YT (Yta/b) | rs1799805 | rev: GGGAGGACTTCTGGGACTTC | ACHE | C__8786419_20 |
| for: CATCAACGCGGGAGACTTCC | c.1057C | |||
| for: CATCAACGCGGGAGACTTCA | c.1057A | |||
| SC (SC1/2) | rs56025238 | rev: CCCTTATATTCCGGCATCAGATC | ERMAP | C__32292873_10 |
| for: CTCTCTCCCTCTGGCCCG | c.169G | |||
| for: CTCTCTCCCTCTGGCCCA | c.169A | |||
| DO (Doa/b) | rs11276 | for: CTCCACATCCCTCCTGAAAG | ART4 | C__2687344_20 |
| rev: GTTGACCTCAACTGCAACCAGTT | c.793A | |||
| rev: GTTGACCTCAACTGCAACCAGTC | c.793G | |||
| CO (Coa/b) | rs28362692 | rev: GCACGGAAGATGCTGATCTGG | AQP1 | C__27536484_20 |
| for: GGGGAACAACCAGACGGC | c.134C | |||
| for: GGGGAACAACCAGACGGT | c.134T | |||
| LAN (+/–) | rs149202834 | rev: AGGTGTACCTGGCTCCTTTC | ABCB6 | C_168788417_10 |
| for: TTTAGCCTGTGGGTGCTGC | c.574C | |||
| for: TTTAGCCTGTGGGTGCTGT | c.574T | |||
| VEL (+/–) | rs566629828 | rev: CCAAAGGCTGCGGTTTGCTG | SMIM1 | see [7] |
| for: GCAGCAGGGACGGAGTCA | c.64–80ins | |||
| for: GCAGCAGGGACGGAGTCC | c.64–80del | |||
| HPA-1a/b | rs5918 | rev: gtgcaatcctctggggact | ITGB3 | C__818008_30 |
| for: gacttacaggccctgcctct | ITGB3*176T | |||
| for: acttacaggccctgcctcc | ITGB3*176C | |||
| HPA-2a/b | rs6065 | rev: gccagcgacgaaaatagagg | GP1BA | C__11442703_10 |
| for: cccccagggctcctgac | GP1BA*482C | |||
| for: cccccagggctcctgat | GP1BA*482T | |||
| HPA-3a/b | rs5911 | rev: accagagagcctgctcactac | ITGA2B | C__3017440_10 |
| for: ggactgggggctgcccat | ITGA2B*2621T | |||
| for: ggactgggggctgcccag | ITGA2B*2621G | |||
| HPA-5a/b | rs1801106 | rev: ggcagtacactatacattcaactc | ITGA2 | C__27862812_10 |
| for: aaggaagagtctacctgtttactatcaaag | ITGA2*1600G | |||
| for: aaggaagagtctacctgtttactatcaaaa | ITGA2*1600A | |||
| HPA-15a/b | rs10455097 | for: cagtattatgaccttatgatgacctattc | CD109 | C__3226894_10 |
| rev: gttacttcaaattcttggtaaatcctgg | CD109*2108C | |||
| rev: gttacttcaaattcttggtaaatcctgt | CD109*2108A | |||
| HNA-1a | rs448740 | for: cacagtggtttcacaatgagaa | FCGR3B*01 | none |
| rs368410676 | rev: atggacttctagctgcacc | |||
| HNA-1b/d | rs527909462 | for: gcctcaatggtacagcgtgctt | FCGR3B*02 | none |
| rs5030738 | rev: ctgtcgttgactgtggcag | |||
| HNA-1b/c | rs527909462 | for: gcctcaatggtacagcgtgctt | FCGR3B*03 | none |
| rs5030738 | rev: ctgtcgttgactgtggcaT | |||
| HNA-2 (+/–) | rs1164364335 | for: catggagaaggtgacctttgag | CD177 | (self-designed assay2) |
| rev: CAACAGTGCTGCAGCCTTT | CD177*787A | |||
| rev: CAACAGCGCTGCAGCTTTA | CD177*787T | |||
| HNA-3a/b | rs2288904 | rev: ctgcatggagcagaggatgg | SLC44A2 | C__11789692_10 |
| for: GAGTGGCTGAGGTGCTTCG | SLC44A2*461G | |||
| for: GGAGTGGCTGAGGTGCTTCA | SLC44A2*461A | |||
| HNA-4a/b | rs1143679 | rev: aaggaggtctgacggtgaag | ITGAM | C__2847895_1_ |
| for: ctcatgcgagcccatccg | ITGAM*230G | |||
| for: ctcatgcgagcccatcca | ITGAM*230A | |||
| HNA-5a/b | rs2230433 | rev: tccaccttgcggaaggagagtc | ITGAL | C___25754090_10 |
| for: atcatcccccacagatccag | ITGAL*2372G | |||
| for: catcccccacagatccac | ITGAL*2372C | |||
*
TaqMan™-PCR assays were commercially available (Life Technologies) under the given assay number
1
self-designed TaqMan™-PCR assay for MN genotyping with forward primer CTCAGTCACCTCGTTCTTAATC, reverse primer GGCAAGAATTCCTCCATAGTAG, FAM-probe CACTGGTGTGGCAA (M), VIC-probe CACTGAGGTGGCAA (N)
2
self-designed TaqMan™-PCR assay for HNA-2 genotyping with forward primer CCCTCAGGACTCACATCAAC, reverse primer CTGAGTGGATGGTGGTCTTC, FAM-probe CAGCCTTTTGTCCC (CD177 wild type), VIC-probe CAGCTTTAGGTCGC (CD177 null variant).