Identification of HLA-C alleles using PCR—single-strand-conformation polymorphism and direct sequencing |
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| Authors: | H Wang K Tokunaga T Akaza K Tadokoro Y Shibata T Juji |
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| Institution: | Department of Research, Japanese Red Cross Central Blood Center;Department of Transfusion Medicine and Immunohematology, Faculty of Medicine, University of Tokyo;Department of Human Genetics, Graduate School of International Health, University of Tokyo, Japan |
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| Abstract: | Alleles encoding HLA-C antigens in Japanese were identified by polymerase chain reaction followed by single strand conformation polymorphism (PCR-SSCP) and nucleotide sequencing analyses. The results showed that at least sixteen different alleles code for eight serologically detectable antigen groups and undetectable blanks. Cwl was mainly encoded by Cw*0102, whereas two split antigens of Cw3, Cw9 and Cw10, were encoded by Cw*0303 and Cw*0304, respectively. Cw4 and Cw6 were encoded by Cw*0401 and Cw*0602, respectively. Seven alleles, Cw*0801, Cw*0803, Cw*1202, Cw*1203, Cw*1402, Cw*1403 and Cw*1502, were found to encode serological HLA-C "blanks" in Japanese. Moreover, errors in the published nucleotide sequences of Cw*0501 and Cw*1201 were corrected. Twenty-one HLA-C alleles were distinguished from each other by means of group-specific PCR amplification followed by the SSCP method developed in the present study. The system using genomic DNAs can be used effectively for identification of new HLA-C alleles. |
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| Keywords: | HLA-C allele PCR-SSCP direct sequencing |
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