Identification of a novel B allele with missense mutation (c.98G > C) in the ABO gene

A novel mutation +5904 C>T of RUNX1 site in the erythroid cell‐specific regulatory element decreases the ABO antigen expression in Chinese population

“…Thirty ABO subtype individuals without any mutation in the coding region or splicing site were collected from volunteer blood donors in Blood Center of Zhejiang Province, China, or from the patients in the hospitals of Hangzhou City, who were screened from 211 ABO subtypes individuals base on our previous studies . All individuals were found with ABO blood group discrepant results between the serologic forward and reverse typing.…”

“…The full coding regions of ABO gene, including exon 1–7 regions, were amplified with three specific primer pairs by polymerase chain reaction. Then, the amplicons were purified with enzyme digestion and directly sequenced according to our previous report . The partial nucleotide sequences of intron 1 covering the +5·8‐kb region were also amplified and directly sequenced.…”

c.518T > C missense mutation in the B glycosyltransferase gene responsible for a weak B variant

“…Reverse typing was determined with A, B, and O cells using tube test by trained staff according to the manufacturer's instructions. The whole coding region of the ABO gene was amplified by polymerase chain reaction and sequenced bidirectionally according to our previous report . The haplotype of Exons 5 through 7 containing the novel mutation of the proband was analyzed by a TOPO TA cloning kit according to the manufacturer's instruction (Invitrogen).…”

“…The haplotype of Exons 5 through 7 containing the novel mutation of the proband was analyzed by a TOPO TA cloning kit according to the manufacturer's instruction (Invitrogen). Plasmid DNA was extracted from each colony and was used as the template for sequencing reactions, as previously described . The transferase activity was assayed by the modified method of Nagai and coworkers…”

“…The whole coding region of the ABO gene was amplified by polymerase chain reaction and sequenced bidirectionally according to our previous report. 2 The haplotype of Exons 5 through 7 containing the novel mutation of the proband was analyzed by a TOPO TA cloning kit according to the manufacturer's instruction (Invitrogen). Plasmid DNA was extracted from each colony and was used as the template for sequencing reactions, as previously described.…”