Abstract:

BACKGROUND: Correlation studies between HLA-DQB1 gene and nasopharyngeal carcinoma are constrained by expensive imported reagents; on the other hand, the genotyping of HLA-DQB1 at exon 2 is ambiguous, and the results are less.
OBJECTIVE: To apply the direct sequencing genotyping method of HLA-DQB1 gene at exons 2 and 3.
METHODS: 286 samples from voluntary healthy donors and 46 samples from nasopharyngeal carcinoma patients were included. Gentra DNA extraction reagent was applied to prepare genomic DNA. 100 samples from healthy donors served as parallel control group for genotyping. We explored the most suitable conditions for PCR amplification and sequencing reaction of HLA-DQB1 allele exons 2 and 3. Post-process HLA-DQB1 gene sequencing parting kit was used for one-way parallel controls.
RESULTS AND CONCLUSION: The results of 100 samples as the parallel control are consistent with those of HLA-SBT genotyping. For 286 healthy samples, 13 HLA-DQB1 alleles were detected and 27 HLA-DQB1 alleles were homozygous in unrelated healthy individuals. Eleventh HLA-DQB1 alleles were detected, and five HLA-DQB1 alleles were homozygous in nasopharyngeal carcinoma patients. The frequency of HLA-DQB1*02:01 in nasopharyngeal carcinoma C patients was higher than that in unrelated healthy individuals; however, the frequency distribution of HLA-DQB1 alleles had not significant differences between the two groups (x² < 3.84, P > 0.05). No significant association between the HLA-DQB1 and nasopharyngeal carcinoma was confirmed. These findings indicate that the method for HLA-DQB1 genotype from the Chinese Han population is easy to operate with stable results and low cost.