The kit is used for in vitro qualitative detection of ABO alleles in human red blood cell genomic DNA, including A102, A201, A205, B101, O01, O02 alleles. This kit cannot be used for blood source screening.

ABO Genotyping Kit (60 tests per kit)

The ABO locus, with 7 exons, is located on human chromosome 9q34, and the A, B, and O genes are highly homologous. The ABO gene is approximately 1060bp and encodes 353 amino acids. The A and B genes encode glycosyltransferases, while the O gene is an invalid gene. There are differences between the B gene and the A gene at the 297bp, 703bp, 796bp, and 803bp four base sites in the cDNA sequence. The difference between the O gene and the A gene is the deletion of the 261bp base G in cDNA. Type A mainly includes subtypes A1 and A2, in addition to A3, Am, and Ax subtypes. Compared with the A1 gene, the A2 gene exhibits a A/T substitution at 467bp of cDNA and a deletion of base C at 1061bp. The main subtypes of O are O1 and O2. Compared with the O1 gene, in addition to the 261bp base G deletion, the O2 gene also has four base substitutions:646T>A, 681G>A, 771C>T, and 829G>A^[1-3]. The ABO blood type system is the most immunogenic system in the human blood type systems, and plays an important role in transfusion medicine, as well as clinical medicine such as organ and stem cell transplantation. Serological techniques for identifying ABO blood type systems can only identify phenotypes and are limited in identifying individuals with abnormal expression and conducting genetic analysis^[4]. The ABO blood type genotyping technology can be combined with serological diagnostic results in transfusion medicine for clinical application. The ABO gene typing results can be used as genetic markers for chimeric detection after stem cell transplantation and the family study on subtypes. This kit designs specific primers for the replacement or deletion of different bases in the ABO gene, including base replacement for 261 base G deletion and intron in the O1 gene, 646T>A and 829G>A in the O2 gene, 803G/C in the B gene, and C deletion at 467 C/T and 1061 positions in the A2 gene. PCR-SSP technology is used for the determination of ABO blood type gene typing in red blood cells.

The designing principle of sequence specific primers polymerase chain reaction (PCR-SSP) is to design primers based on the differences in a certain base of an allele. Specific primers only amplify the corresponding allele and do not amplify other alleles. Therefore, the presence or absence of amplification products is the basis for identifying specific alleles The specific PCR products can be detected by agarose gel electrophoresis. Specific primers, pointing at ABO related gene mutations, deletions, insertions and other information, were designed according to the sequence information published in Genbank database of NCBI.
The internal reference primers were designed based on the sequence of the conserved regions of human genes. Internal reference primer pairs are used to check the amplification effectiveness of the reaction systems. They are coated with specific primers in each reaction well to amplify together.