Molecular Analysis of HLA-C Using Polymerase Chain Reaction-Sequence Specific Primers

  • Lee, Kyung-Ok (Seoul Clinical Laboratories, Seoul Medical Science Institute) ;
  • Hong, Sung-Hoi (Seoul Clinical Laboratories, Seoul Medical Science Institute) ;
  • Kim, Min-Jung (Department of Chemistry, College of Science, Kon-Kuk University) ;
  • Park, Taek-Kyu (Department of Chemistry, College of Science, Kon-Kuk University) ;
  • Kim, Yoon-Jung (Seoul Clinical Laboratories, Seoul Medical Science Institute) ;
  • Lee, Kyu-Pum (Seoul Clinical Laboratories, Seoul Medical Science Institute)
  • Received : 1996.10.16
  • Published : 1997.01.31

Abstract

Of all HLA class I molecules, HLA-C gene products are most poorly understood because they express at a low level on the cell surface compared to HLA-A and -B. In order to identify serologically detectable and undetectable HLA-C antigens, we have established a DNA-based tissue typing method for the HLA-C locus by PCR-SSP (polymerase chain reaction-sequence specific primers). Genomic DNA prepared from Iymphoblastoid 21 B-cell lines and 120 Korean individuals by proteinase K digestion and pheno/chloroform extractions have been typed by PCR-SSP (23 primer mixes were used). The PCR-SSP results of control cell lines were discrepant from serology in 1 case among 21 cases: Cw6 which was negative by serology but positive by PCR-SSP (cell line: MANIKA). Twenty four HLA-Cw "blank" antigens among fifty Korean individuals were completely determined by PCR-SSP DNA typing. HLA-Cw*0101 (15.3%), Cw*1401 (12.3%) and Cw*0701 (11.7%) alleles were frequently found in 120 Korean individual samples. In conclusion. the high level of discrimination for HLA-C alleles may prove useful and informative in the study of transplant survival, and identify the importance of allelic differences, not readily detectable by serology, on host and donor compatibility.

Keywords

DNA typing;HLA-C;polymerase chain reaction-sequence specific primers

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