Analysis of HLA DQαusing Polymerase Chain Reaction and Allele Specific Oligonucleotide Method in Korean Insulin Dependent Diabetes Mellitus Patients

다국어 초록 (Multilingual Abstract)

Genetic predispostition to the autoimmune disease insulin dependant diabetes mellitus is determined, in part, by a gene within the major histocompatibility complex (MHC) on human chromosome 6.
After amplification of exon 2 HLA-DQα chain gene with polymerase chain reaction (PCR) procedure, we used non-radioactive HLA-DQα ASO probes which labelled with horseradish peroxidase (HRP) coupled to the 5’ end of the oligonucleotide for the HLA-DQα typing.
The results were as follows:
1. The first domain elf the DQα chain gene was amplified from minimum 1 ng of genomic DNA with use of the PCR. And from 1 ㎍ DNA samples, all the samples were successfully amplified which have a product of approximately 242 bp.
2. On oligonulcleotide dot blot analysis of study subjects and standard control cell lines, TAB (HLA-DQAI homozygosity) cell line was hybridized to RH 83 probe only and DKB (HLA-DQA3 homozygosity) cell line to GH 67 probe and LUY (HLA-DQA4 homozygosity) cell line to GH 66. According to these results, we accurately determined HLA-DQα subtypes in study subjects.
3. Frequency of DQA3 allele in Korean was high and DQA2 allele was low compared to other populations. In IDDM patients, DQA3 allele w as significantly increased (relative risk 6.6, P=0.03) compared with controls. Conversely the frequencies of DQA1 allele was decreased among patients (relative risk 0.25, P=0.02).
These results suggest that HLA-DQA3 allele is positively associated with IDDM as in other populations and HLA-DQA1 allele is negatively associated with IDDM in Korean population.