The Discovery And Confirmation Research Of HLA-B~*48:22 Novel Allele

OBJECTIVE Our laboratory assumed the HLA genotyping work of voluntary donors from Tianjin Registry of the China Marrow Donor Program. The HLA genotyping result of one sample could not be confirmed by direct sequencing technology. Gene cloning and sequencing were completed to isolate the single unknown allele sequence to identify the potential novel HLA allele that the sample might carry.METHODS HLA PCR-SSO genotyping, PCR-SBT genoyping,cloning and sequencing technology were used to study the potential novel HLA allele in this project with commercial reagents. Sequence alignments with IMGT professional database were completed after single HLA allele sequence was obtained. The unknown sequence was registered in GenBank to accquire accession number, and then submitted to the WHO HLA Nomenclature Committee.RESULTS (1) Cloning and sequencing technology was successfully applied to establish a research method to separate single HLA allele.(2) Separate sequence of the unknown HLA allele was accquired and sequence alignment was completed in the MGT professional database.Three base substitution appeared in the HLA-B second exon (Exon2) sequence compared with the nearest HLA-B*48:01:01 sequence. The unknown sequence was submitted to WHO HLA Nomenclature Committee.CONCLUSIONS Sequence of the unknown HLA allele was registered in GenBank.The accession number is GU570429. The unknown Sequence has been officially named HLA-B *48:22 by WHO HLA Nomenclature Committee on January 31,2010.