Xu H.,Blood Group Laboratory |
Ye S.-H.,Blood Group Laboratory |
Wang B.-Y.,Xian Jiaotong University |
Liu M.-L.,Blood Group Laboratory |
And 2 more authors.
Journal of Xi'an Jiaotong University (Medical Sciences) | Year: 2010
Objective: To establish a reliable PCR method for RHD genotypes research. Methods: Based on the characteristics of RHD gene, the specific primers were designed for different RHD genotypes to establish the multiplex PCR and compare it with commercially supplied RHD genotyping kit. Results: The multiplex PCR utilized for RHD genotypes was established. The results of RHD genotypes were the same between the multiplex PCR method and commercially supplied kit. Conclusion: The method of the multiplex PCR can be utilized for the study of RHD genotypes because it is convenient, accurate and economical.
Cai X.,Shanghai Institute of Blood Transfusion |
Jin S.,Blood Group Laboratory |
Liu X.,Blood Group Laboratory |
Fan L.,Blood Group Laboratory |
And 6 more authors.
Transfusion | Year: 2013
BACKGROUND: Identifying genetic variants of the ABO gene may reveal new biologic mechanisms underlying variant phenotypes of the ABO blood group. We report the molecular genetic analysis of 322 apparently unrelated ABO subgroup individuals in an estimated 2.1 million donors. STUDY DESIGN AND METHODS: We performed phenotype investigations by serology studies, analyzed the DNA sequence of the ABO gene by direct sequencing or sequencing after cloning, and evaluated promoter activity by reporter assays. RESULTS: In 62 rare ABO alleles, we identified 29 novel ABO subgroup alleles in 43 apparently unrelated subgroup individuals and their four available pedigrees. Of these alleles, one was a deletion-mutation allele, four were hybrid alleles, and 24 were point-mutation alleles. Most of the point mutations were detected in Exons 6 to 7, while several others were also detected in Exons 1 to 5 or splicing regions. One ABO promoter mutation, -35 to -18 del, was found and verified to reduce promoter activity, as determined by dual luciferase assays. Two mutations, 7G>T and 52C>T, carrying the premature terminal codons E3X and R18X in the 5'-region, were found to be associated with the very weak ABO subgroups "Ael" and "Bel." CONCLUSION: Twenty-nine ABO subgroup alleles were newly linked to different kinds of ABO variations. We provide the first evidence that promoter abnormality is involved in the formation of weak ABO phenotypes. We also described the first naturally occurring ABO alleles with premature terminal codons in the 5'-region that led to Ael and Bel phenotypes.