Isoelectric focusing subtypes of HLA‐A can be defined by oligonucleotide typing Journal Article
| Authors: | Oh, S. H.; Fleischhauer, K.; Yang, S. Y. |
| Article Title: | Isoelectric focusing subtypes of HLA‐A can be defined by oligonucleotide typing |
| Abstract: | This study describes a simple and direct method for sequence‐specific oligonucleotide probe (SSOP) typing of the A locus of HLA class I genes. Genomic DNA from a panel of over 200 cells which have been characterized by the methods of serology and isoelectric focusing (IEF) for the HLA class I antigens was used for locus‐specific PCR amplification of HLA‐A sequences. Dot blot hybridization of the amplified products was performed with 28 SSOPs derived from hypervariable regions in exon 2 and 3. Co‐amplification of three alleles of HLA‐H pseudogene in apparent linkage disequilibrium with HLA‐A2 and A10 was observed but did not interfere with the typing of HLA‐A alleles. Using short SSOPs (15 nucleotides each) in single temperature tetramethylammonium chloride (TMAC) hybridization and washing steps, 30 IEF‐definable isotypes of HLA‐A antigens could be unambiguously defined by their hybridization patterns. Moreover, comparison of the typing results with available nucleotide sequences of HLA‐A alleles showed that the conditions used allowed faithful detection of single codon mismatches between probe and template. Thus, these alleles can be identified by their unique hybridization patterns generated by the SSOPs. Nucleotide sequence analysis of any new HLA‐A allele will further permit its rapid and unambiguous characterization by SSOP typing. 1993 Blackwell Munksgaard |
| Keywords: | human cell; exon; comparative study; sensitivity and specificity; polymerase chain reaction; allele; gene amplification; cell line; gene locus; gene frequency; amino acid sequence; molecular sequence data; sequence alignment; nucleotide sequence; hla antigen class 1; base sequence; codon; sequence homology; sequence homology, nucleic acid; oligonucleotide; serology; hla-a antigens; normal human; cell dna; histocompatibility testing; histocompatibility test; dna template; consensus sequence; pseudogene; pseudogenes; gene linkage disequilibrium; dna hybridization; isoelectric focusing; human; priority journal; article; support, u.s. gov't, p.h.s.; ief; dot hybridization; dna probes, hla; hla‐a; ssop typing; tetramethylammonium chloride |
| Journal Title: | Tissue Antigens |
| Volume: | 41 |
| Issue: | 3 |
| ISSN: | 0001-2815 |
| Publisher: | Wiley-Blackwell Publishing, Inc. |
| Date Published: | 1993-03-01 |
| Start Page: | 135 |
| End Page: | 142 |
| Language: | English |
| DOI: | 10.1111/j.1399-0039.1993.tb01992.x |
| PUBMED: | 8316944 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 1 March 2019 -- Source: Scopus |
