Extended Donor Typing by Pooled Capillary Electrophoresis: Impact in a Routine Setting

Area of Science:

• Molecular Biology
• Immunogenetics
• Transfusion Medicine

Background:

• Polymerase chain reaction (PCR) with sequence-specific priming and fluorescent primers is a standard DNA typing technique.
• Adapting this method for medium-throughput donor typing was the primary objective.

Purpose of the Study:

• To develop and validate a multiplex allele-specific PCR method for extended donor DNA typing.
• To assess the cost-effectiveness and routine performance of the developed method.

Main Methods:

• Eight multiplex allele-specific PCR assays with fluorescent primers were designed for common blood group antigens.
• Capillary electrophoresis was used for analysis, with fluorescent color discriminating alleles and product length encoding donor/polymorphism.
• Sequencing was employed for discrepant samples, and statistical analysis evaluated new allele associations.

Main Results:

• The method achieved 99.96% prediction accuracy on validation with 312 samples.
• Consumable costs per donor were below EUR 2.00, with 92.2% successful prediction in routine use.
• Eleven novel alleles were identified, and a significant portion of red blood cell units were from donors with predicted antigen status, ensuring supply of rare phenotypes.

Conclusions:

• Pooled capillary electrophoresis is a viable alternative for extended donor DNA typing in a medium-throughput setting.
• Establishing a large cohort of typed donors enhances transfusion support for patients, particularly those requiring rare blood types.