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6,7 Since data for the haplotype DR4�CDQ8 and alleles DRB1*0401, DQA1*0301, and DQB1*0302 is not available for the children of East Azerbaijanpopulation, the current study aims to evaluate the genetic association between this haplotype and DM-1A risk. Materials and Methods Subjects Eighty unrelated DM-1A patients were recruited from northwest Iran. DM-1A was diagnosed by the consultant endocrinologist according to clinical features and results of laboratory tests. The allele frequency of the HLA class II DR4�CDQ8 haplotype was determined in the DM-1A patients. In addition, 80 control subjects with normal fasting/random blood glucose levels and no family history of DM-1A or other autoimmune diseases were included in the study (Table 1). Table 1 Characteristic of DM-1A patients and normal subjects HLA genotyping Temsirolimus ic50 Total genomic DNA was extracted from the peripheral PD173074 nmr blood of study participants using the salting-out method, and used for polymerase chain reaction (PCR) analysis. PCR-Sequence-Specific Priming (PCR-SSP) technique was used for genotyping of DRB1*0401-DQB1*0302-DQA1*0301 haplotype, as describedby IonescuTirgoviste et al.8 The PCR products were analyzed by agarose gel electrophoresis using a 1% (w/v) agarose gel, and visualized by staining with Simply Blue safe stain (Fermentas). Data analysis Data was analyzed using STATA11 statistical software package (STATA Corporation, Texas). For preliminary descriptive statistics, variant analyses were done using chi-square test and calculation of odd ratios (ORs) with 95% confidence interval (CI). For analyses of haplotype effects, the logistic regression modules developed for STATA were used. This procedure generated tabulated values detailing frequency probability of various alleles along with adjusted ORs for minor alleles compared to the reference major allele. MRIP P value and 95% CI were also reported. P value