Journal article
A DNA-based detection and screening system for identifying HLA class I expression variants by sequence-specific primers
Tissue Antigens, Vol.53(5), pp.498-506
1999
Abstract
Molecular methods are now commonplace for HLA typing and they have replaced traditional serological methods in many histocompatibility laboratories. A consequence of reliance on molecular methods using primers or probes based on existing sequence information is that unsequenced or partially-sequenced null, or low expressed variants are not discriminated from expressed alleles. Failure to identify null alleles might have deleterious implications for allogeneic transplants. Expression variants may be classified into two categories: unique mutations and repeat mutations. For example, the alleles A*0303N, A*2409N, and B*1526N have apparently unique mutations. In contrast, repeat mutations may occur frequently at points where unusual nucleotide sequences make accurate DNA replication by DNA polymerases difficult. One example is between nucleotide positions 621–627, where HLA class I alleles may exhibit between three and seven consecutive cytosine residues. Incorrect insertion of an extra cytosine in this region is the cause of expression failure in A*2411N and A*0104N alleles. We hypothesise that insertion of an extra cytosine into the cytosine island between nucleotide positions 621–627 is likely to recur not only in other HLA-A alleles but also in HLA-B and even HLA-C alleles. We describe here a polymerase chain reaction using sequence-specific primers (PCR-SSP) system that can not only detect all previously-sequenced HLA class I expression variants but can also screen for mutations between positions 621–627 in HLA-A, B or C alleles which may give rise to potentially null alleles. Overall, in this study HLA class I expression variants were identified in 5 of 931 tested samples (0.53%).
Details
- Title
- A DNA-based detection and screening system for identifying HLA class I expression variants by sequence-specific primers
- Authors/Creators
- M. Bunce (Author/Creator) - Churchill HospitalJ. Procter (Author/Creator) - Churchill HospitalK.I. Welsh (Author/Creator) - Churchill Hospital
- Publication Details
- Tissue Antigens, Vol.53(5), pp.498-506
- Publisher
- Blackwell Publishing
- Identifiers
- 991005542907007891
- Copyright
- 1999 Munksgaard
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
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- Citation topics
- 1 Clinical & Life Sciences
- 1.6 Immunology
- 1.6.607 MHC Diversity
- Web Of Science research areas
- Cell Biology
- Immunology
- Pathology
- ESI research areas
- Molecular Biology & Genetics