Objective To evaluate the status of eight RHD specific exons in 131 Han Chinese blood donors who were classified as RhD-negative by serological methods and explore the genomic structure of RHD gene among the Han Chine...Objective To evaluate the status of eight RHD specific exons in 131 Han Chinese blood donors who were classified as RhD-negative by serological methods and explore the genomic structure of RHD gene among the Han Chinese. The Rh blood group system has the highest prevalence of polymorphisms among human blood group systems and is clinically significant in transfusion medicine. The Rh antigens are expressed on polypeptides encoded by two highly homologous genes,RHD and RHCE. Recent molecular studies have shown that the RhD-negative trait could be generated by multiple genetic mechanisms and is ethnic group-dependent. Methods The polymerase chain reaction using-sequence specific primers (PCR-SSP) was used to amplify exons 2,3,4,5,6,7,9 and 10 of RHD gene and exons 1,2 and 5 of RHCE gene,as well as intron 4 in each of them.Results The 131 cases of RhD-negative phenotypes consisted of 60 ccee,58 Ccee,5 ccEe,5 CcEe and 3 CCee. Among them,83 with the Rh ccee or ccEe phenotypes (63.4%) lacked the eight RHD exons indicated above,while 26 cases with the Rh Ccee,CCee,CcEe phenotypes (19.9%) had all the RHD exons examined. Twenty-two individuals with the Ccee,CCee,CcEe phenotypes (16.8%) carried at least one RHD exon. The phenotypes of the RhD negative individuals carrying the RHD gene were Rh CC or Cc,but not cc. Conclusions Three classes of RhD-negative polymorphisms among a population of Han Chinese were observed. Antigen association analysis suggested the existence of a novel class of RhD-negative associated haplotype in Han Chinese. This haplotype consisted of a normal RHCE allele and a nonfunctional RHD gene. It may be beneficial to redefine the RhD-negative blood group among Chinese populations upon clarification of the mechanisms of RHD gene expression and RhD antigen immunization.展开更多
In order to investigate the association of genotypes of HLA-DRB1 and HLA-DQB1 alleles with the genetic susceptibility of chronic urticaria (CU), genotypes of HLA-DRB1 and HLA-DQB1 genes were detected by polymerase cha...In order to investigate the association of genotypes of HLA-DRB1 and HLA-DQB1 alleles with the genetic susceptibility of chronic urticaria (CU), genotypes of HLA-DRB1 and HLA-DQB1 genes were detected by polymerase chain reactions with sequence-specific primers (PCR-SSP) in 42 patients with CU (19 men and 23 women, mean age 30.67±12.45 y old as well as 193 racially matched healthy persons in ethnic Han from Hubei provinece. Gene frequencies of HLA-DRB1*12, *0901 (RR=3.11, χ2=7.579, P=0.006; RR=2.47, χ2=5.684, P=0.017) were significantly increased in CU patients as compared with that in healthy people. Gene frequencies of HLA-DQB1*05 (RR=0.26, χ2=6.683, P=0.01) were significantly decreased in CU patients. It was suggested that CU was found strongly associated with HLA-DRB1*12, *0901 and HLA-DQB1*05, the former might be the genetic markers for susceptibility to CU, but the latter might play a resistive role.展开更多
Background Molecular testing is more precise compared to serology and has been widely used in genotyping blood group antigens. Single nucleotide polymorphisms （SNPs） of blood group antigens can be determined by the ...Background Molecular testing is more precise compared to serology and has been widely used in genotyping blood group antigens. Single nucleotide polymorphisms （SNPs） of blood group antigens can be determined by the polymerase chain reaction with sequence specific priming （PCR-SSP） assay. Commercial high-throughput platforms can be expensive and are not approved in China. The genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE blood group antigens in Jiangsu province were unknown. The aim of this study is sought to detect the genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE antigens in Jiangsu Chinese Hart using molecular methods with laboratory developed tests. Methods DNA was extracted from EDTA-anticoagulated blood samples of 146 voluntary blood donors collected randomly within one month. Standard serologic assay for red blood cell antigens were also performed except the Scianna blood group antigens. PCR-SSP was designed to work under one PCR program to identify the following SNPs： JK1/JK2, KEL 1/KEL2, FYA/FYB, SC1/SC2, C/c and E/e. Results Serologic antigen results were identical to the phenotypes that were predicted from genotyping results. The allele frequencies for Jk^＊01 and Jk^＊02 were 0.51 and 0.49, respectively; for Fy^＊A and Fy^＊B 0.94 and 0.06; for RHCE^＊C and RHCE^＊c 0.68 and 0.32; and for RHCE^＊E and RHCE^＊e 0.28 and 0.72. Among 146 blood donors, all were KEL^＊02/ KEL^＊02 and SC^＊01/SC^＊01, indicating allele frequencies for KEL^＊02 and SC^＊01 close to 1.00. Conclusions The use of PCR-SSP working under the same condition for testing multiple antigens at the same time is practical. This approach can be effective and cost-efficient for small-scale laboratories and in developing counties. These molecular tests can be also used for identifying rare blood types.展开更多
基金ThisprojectwassupportedbyagrantfromShandongProvincialHealthBureauFoundationforYoungScientists (No .1999C3 4)
文摘Objective To evaluate the status of eight RHD specific exons in 131 Han Chinese blood donors who were classified as RhD-negative by serological methods and explore the genomic structure of RHD gene among the Han Chinese. The Rh blood group system has the highest prevalence of polymorphisms among human blood group systems and is clinically significant in transfusion medicine. The Rh antigens are expressed on polypeptides encoded by two highly homologous genes,RHD and RHCE. Recent molecular studies have shown that the RhD-negative trait could be generated by multiple genetic mechanisms and is ethnic group-dependent. Methods The polymerase chain reaction using-sequence specific primers (PCR-SSP) was used to amplify exons 2,3,4,5,6,7,9 and 10 of RHD gene and exons 1,2 and 5 of RHCE gene,as well as intron 4 in each of them.Results The 131 cases of RhD-negative phenotypes consisted of 60 ccee,58 Ccee,5 ccEe,5 CcEe and 3 CCee. Among them,83 with the Rh ccee or ccEe phenotypes (63.4%) lacked the eight RHD exons indicated above,while 26 cases with the Rh Ccee,CCee,CcEe phenotypes (19.9%) had all the RHD exons examined. Twenty-two individuals with the Ccee,CCee,CcEe phenotypes (16.8%) carried at least one RHD exon. The phenotypes of the RhD negative individuals carrying the RHD gene were Rh CC or Cc,but not cc. Conclusions Three classes of RhD-negative polymorphisms among a population of Han Chinese were observed. Antigen association analysis suggested the existence of a novel class of RhD-negative associated haplotype in Han Chinese. This haplotype consisted of a normal RHCE allele and a nonfunctional RHD gene. It may be beneficial to redefine the RhD-negative blood group among Chinese populations upon clarification of the mechanisms of RHD gene expression and RhD antigen immunization.
文摘In order to investigate the association of genotypes of HLA-DRB1 and HLA-DQB1 alleles with the genetic susceptibility of chronic urticaria (CU), genotypes of HLA-DRB1 and HLA-DQB1 genes were detected by polymerase chain reactions with sequence-specific primers (PCR-SSP) in 42 patients with CU (19 men and 23 women, mean age 30.67±12.45 y old as well as 193 racially matched healthy persons in ethnic Han from Hubei provinece. Gene frequencies of HLA-DRB1*12, *0901 (RR=3.11, χ2=7.579, P=0.006; RR=2.47, χ2=5.684, P=0.017) were significantly increased in CU patients as compared with that in healthy people. Gene frequencies of HLA-DQB1*05 (RR=0.26, χ2=6.683, P=0.01) were significantly decreased in CU patients. It was suggested that CU was found strongly associated with HLA-DRB1*12, *0901 and HLA-DQB1*05, the former might be the genetic markers for susceptibility to CU, but the latter might play a resistive role.
文摘Background Molecular testing is more precise compared to serology and has been widely used in genotyping blood group antigens. Single nucleotide polymorphisms （SNPs） of blood group antigens can be determined by the polymerase chain reaction with sequence specific priming （PCR-SSP） assay. Commercial high-throughput platforms can be expensive and are not approved in China. The genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE blood group antigens in Jiangsu province were unknown. The aim of this study is sought to detect the genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE antigens in Jiangsu Chinese Hart using molecular methods with laboratory developed tests. Methods DNA was extracted from EDTA-anticoagulated blood samples of 146 voluntary blood donors collected randomly within one month. Standard serologic assay for red blood cell antigens were also performed except the Scianna blood group antigens. PCR-SSP was designed to work under one PCR program to identify the following SNPs： JK1/JK2, KEL 1/KEL2, FYA/FYB, SC1/SC2, C/c and E/e. Results Serologic antigen results were identical to the phenotypes that were predicted from genotyping results. The allele frequencies for Jk^＊01 and Jk^＊02 were 0.51 and 0.49, respectively; for Fy^＊A and Fy^＊B 0.94 and 0.06; for RHCE^＊C and RHCE^＊c 0.68 and 0.32; and for RHCE^＊E and RHCE^＊e 0.28 and 0.72. Among 146 blood donors, all were KEL^＊02/ KEL^＊02 and SC^＊01/SC^＊01, indicating allele frequencies for KEL^＊02 and SC^＊01 close to 1.00. Conclusions The use of PCR-SSP working under the same condition for testing multiple antigens at the same time is practical. This approach can be effective and cost-efficient for small-scale laboratories and in developing counties. These molecular tests can be also used for identifying rare blood types.
