Rapid rare ABO blood typing using a single PCR based on a multiplex SNaPshot reaction

Area of Science:

• Genetics
• Hematology
• Molecular Biology

Background:

• ABO subgroups require specialized testing when discrepancies arise in blood grouping.
• Current serological methods for identifying ABO subgroups are laborious and complex.
• There is a need for a reliable and affordable method to assess ABO subgroups.

Purpose of the Study:

• To develop and validate a multiplex SNaPshot-based assay for determining rare A and B blood group subgroups.
• To design specific primer probes for identifying known rare ABO subtypes in the Taiwanese population.

Main Methods:

• Designed multiplex SNaPshot assays with specific primer probes targeting single nucleotide polymorphism (SNP) sites.
• Validated the accuracy and reproducibility of the SNaPshot panel using various ABO subtype samples.

Main Results:

• Successfully designed primer probes for 8 SNP sites for A phenotype and 6 SNP sites for B phenotype.
• Enabled SNaPshot analysis for defining various alleles of blood group A (A1, A2, A3, Am, Ael) and blood group B (B1, B3, Bw, Bel).

Conclusions:

• SNaPshot analysis provides a viable method for typing known rare A and B subgroups in reference laboratories, bypassing DNA cloning and sequencing.
• This genotyping approach can facilitate the creation of blood donor databases and assist in identifying fetal-maternal ABO incompatibility.