HLA Design
HLA Sequencing-based Typing from ABBOTT Molecular
AlleleSEQR kits have one single tube amplification upfront followed by 3— 6 sequencing reactions of the relevant exons.
AlleleSEQR kits have one single tube amplification upfront followed by 3— 6 sequencing reactions of the relevant exons.
Class 1
Sequencing-based Typing
Ambiguity Resolution
Following initial sequencing there is the possibility of
obtaining an ambiguous typing result. For HLA-A and -B the
following group specific amplification (GSA) packs can be
used to resolve most common ambiguities.
The table describes the broad specificities of the five HLA-B Group Specific PCR mixes (BG-1 to BG-5) based on exon 1 polymorphism. Specificity of the mixes for HLA-B alleles not sequenced in exon 1, have not been determined. The PCR mixes allow sequencing of exons 2 and 3 only. PCR amplicons produced with these reagent cannot be sequenced for exon 4.
Sequencing-based Typing
| AlleleSEQR | |
| HLA-A | Single tube amplification followed by 6 sequencing reactions for exons 2, 3, and 4 |
| HLA-B | Single tube amplification followed by 6 sequencing reactions for exons 2, 3, and 4 |
| HLA-C | Single tube amplification followed by 4 sequencing reactions for exons 2 and 3 |
Ambiguity Resolution
Following initial sequencing there is the possibility of
obtaining an ambiguous typing result. For HLA-A and -B the
following group specific amplification (GSA) packs can be
used to resolve most common ambiguities.
| HLA-A2 GSA | HLA-A2 group-specific PCR pre-mix will allow most common ambiguities to be resolved (e.g. A*0201/A*2402, A*0212/A*2413) |
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| HLA-B GSA | 5 group-specific amplifications will allow most common ambiguities to be resolved |
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The table describes the broad specificities of the five HLA-B Group Specific PCR mixes (BG-1 to BG-5) based on exon 1 polymorphism. Specificity of the mixes for HLA-B alleles not sequenced in exon 1, have not been determined. The PCR mixes allow sequencing of exons 2 and 3 only. PCR amplicons produced with these reagent cannot be sequenced for exon 4.
Class II
Sequencing-based Typing
Ambiguity Resolution
The codon 86 sequence is useful for resolving the majority of DRB1 ambiguities, for those not resolved by this method there are two strategies to choose from:
Sequencing-based Typing
| AlleleSEQR | |
HLA-DRB1 | Single tube amplification of exon 2 followed by 3 sequencing reactions (fwd and rev and codon 86) |
HLA-DQB1 | Single tube amplification of exon 2 and exon 3 fol-lowed by 3 sequencing reactions (fwd and rev of exon 2 and one direction of exon 3 to resolve |
DQB1*0201/ 0202 or 0301/0309) | |
HLA-DPB1 | Single tube amplification of exon 2 followed by 3 sequencing reactions (fwd and rev and codon 85) |
Ambiguity Resolution
The codon 86 sequence is useful for resolving the majority of DRB1 ambiguities, for those not resolved by this method there are two strategies to choose from:
| AlleleSEQR | |
HLA-DRB1 GSA | Contains 8 allele group specific PCR pre-mixes |
allow specific amplification of DR groups 1, 2, | |
3/11/6, 4, 7, 8/12, 9 and 10. | |
HLA-DRB1 GSSP | Contains 6 allele group specific sequencing mixes |
to allow specific amplification of DR groups 1, 2, | |
3/11/6, 4, 7 and 8/12. | |
CE Marked Not available in United States
CENAUS
CE Marked
