1.Development of oligoneucleotide arrays for HLA-DR53-associated group genotyping.
Jia-quan XIAO ; Cheng-tao LI ; Jian-ming TAN ; Min-hua KANG ; Yao LI
Chinese Journal of Surgery 2003;41(3):225-227
OBJECTIVETo develop a method performed on an oligonucleotide array for HLA-DR53 group genotyping.
METHODSAccording to the specific allelic frequency and sequence of HLA-DRB loci in Chinese Han population, HLA-DR53 group typing probes which were immobilized on a glass supports were synthesized. A pair of group-special primers labeled by the Cy5-dCTP were designed, and the primers were used in the PCR, thus the PCR products were labeled with Cy5. The labeled PCR products were hybridized with array. The signals were scanned by scanner and analyzed by image software. The typing results were confirmed by standard DNA and PCR-SSO. One hundred and eleven samples were typed by this array.
RESULTSThere were 72 HLA-DR53 group loci typed by oligonucleotide array. Among them, 34 loci were DR9, 25 were DR4, and 13 were DR7. No false positive or false negative typing results were observed. The specificity and reproducibility were 100% and the overall time of genotyping was 5 hours.
CONCLUSIONThe oligonucleotide array technique is a precise, rapid molecular method for HLA-DR53 genotyping, suited for clinical practice.
Genotype ; HLA-DR Antigens ; genetics ; HLA-DRB4 Chains ; Humans ; Oligonucleotide Array Sequence Analysis ; Reproducibility of Results ; Sensitivity and Specificity
2.The relationship between nonresponse to hepatitis B vaccine and HLA genotype/haplotype.
Mingyue LI ; Rongcheng LI ; Shangzhi HUANG ; Jian GONG ; Xianjia ZENG ; Yanping LI ; Ming LU ; Hui LI
Chinese Journal of Preventive Medicine 2002;36(3):180-183
OBJECTIVETo study the relationship between the nonresponse to hepatitis B vaccine and HLA genotype/heplotype in Chinese population and provide the evidence for explaining the genetic mechanism of this nonresponse.
METHODSOur research focused on the relationship between nonresponse to Hepatitis B vaccine and HLA-DRB1, DRB3, DRB4, DRB5 and DQB1 genotype/haplotype in Chinese population, collected from a community in Guangxi Zhuang Autonomous Region. The group specific amplification was employed to characterize 107 individuals' genotype and haplotype of HLA clusters. Different models statistics such as relative risk test, correlation test and linkage disequilibrium analysis were used to analyze the data.
RESULTSThe results showed that there is a linkage disequilibrium between nonresponse to Hepatitis B vaccine and HLA haplotype DR4, 1122 (DRB1 * 0401- 22, 1122)-DR53 (DRB4 * 0101101, 0102/3)-DQB4 (DQB1 * 04).
CONCLUSIONIn Chinese population, nonresponse to hepatitis B vaccine is highly associated with special HLA haplotye.
Asian Continental Ancestry Group ; genetics ; China ; Genotype ; HLA-DQ Antigens ; classification ; genetics ; HLA-DQ beta-Chains ; HLA-DR Antigens ; classification ; genetics ; HLA-DRB1 Chains ; HLA-DRB3 Chains ; HLA-DRB4 Chains ; HLA-DRB5 Chains ; Haplotypes ; Hepatitis B ; genetics ; immunology ; prevention & control ; Hepatitis B Vaccines ; immunology ; Humans ; Linkage Disequilibrium