ABSTRACT
【Objective】 To study the distribution and haplotype polymorphism of HLA-A, -B, -C, -DRB1, -DQB1 alleles in Anhui Han population. 【Methods】 The HLA-A, -B, -C, -DRB1 and -DQB1 genotyping of 3 169 random unrelated stem cell donors was performed by PCR-SBT. The allele frequency, haplotype frequency and linkage imbalance parameters were calculated by counting method, maximum expectation algorithm and PyPop software. 【Results】 A total of 411 HLA alleles were detected in the population, of which 67, 143, 65, 75 and 64 alleles were detected for HLA-A, -B, -C, -DRB1 and -DQB1, respectively. The alleles with frequency >0.1 were HLA-A*11∶01, A*11∶01, A*24∶02, A*02∶01, C*01∶02, C*07∶02, C*06∶02, DRB1*09∶01, DRB1*15∶01, DRB1*07∶01, DQB1* 03∶01, DQB1* 03∶03, and DQB1*02∶01. 1426 HLA-A~HLA-B, 1 772 HLA-B~HLA-DRB1, 798 HLA-B~HLA-C, and 446 HLA-DRB1~HLA-DQB1 haplotypes were detected. The haplotypes showed linkage imbalance, and 19 of them showed strong linkage imbalance (RLD>0.80). 【Conclusion】 The frequency and haplotype distribution of HLA-A, -B, -C, -DRB1 and -DQB1 alleles in Anhui Han population were obtained. The distribution of those alleles and haplotypes have their own characteristics.
ABSTRACT
Objective:To research the distribution features of HLA-A,B and DRB1 high resolution alleles and haplotypes in Heilongjiang population.Methods:PCR-SBT methods was applied for HLA-A,B,DRB1 high resolution genotyping of 13 670 unrelated and healthy donors in region of Heilongjiang,and haplotype frequencies were calculated by counting and maximum likelihood method. Results:A total of 286 HLA alleles were observed and the most frequent alleles(>0.1)were A*02∶01,A*24∶02,A*11∶01,DR*07∶01,DR*09∶01 and DR*15∶01.Among 1 087 kinds of HLA-A-B haplotype ,there were 22 kinds frequency higher than 0.01,and 267 kinds with statistically significant and positive linkage disequilibrium(ALD>0,HF≥1.09×10-4,χ2>3.84).Moreover,among 1 329 kinds of HLA-B-DRB1 haplotype,there were 19 haplotypes frequency higher than 0.01,and 357 kinds with statistically significant and positive linkage disequilibrium.1 348 kinds of A-B-DR Haplotype were informative with frequency≥1.66 ×10-4 in 4 428 haplotypes, and a total of 17 kinds of A-B-DR haplotype frequency higher than 0.005.Conclusion: Get the distribution features of HLA high resolution allele and haplotype in Heilongjiang population, and associated genetic parameters, Distribution of alleles and haplotypes close to northern Han population,but have their own distribution.
ABSTRACT
Ahigh-resolutionmethodforhumanleukocyteantigen-B(HLA-B)genotypingwasestablished based on the optimized polymerase chain reaction, cloning and sequencing technology. The exon2 and exon3 of HLA-B gene were amplified with primers based on the HLA-B gene sequence. These produced heterozygous alleles were effectively cloned into plasmid DNA based on the principle of plasmid incompatibility, and were followed by bacterial culture. Then Sanger sequencing was carried out and after analyzing the result by software ClustalX2 and IMTG/HLA database comparison, the HLA-B genotype of the samples was achieved. Seven clinical samples were detected, and the results were consistent with those of PCR-SBT genotyping method. The method was cost-effective, high-resolution and it did not require technical software. The use of universal primers simplified the cumbersome design and optimization process of specific primers.