Multiplex Lateral Flow Assay for Rapid Visual Blood Group Genotyping.

Area of Science:

• Immunohematology
• Molecular Diagnostics
• Genetics

Background:

• Conventional blood group phenotyping methods are time-consuming and unsuitable for emergency situations.
• Molecular typing offers genotype-based phenotype deduction but often involves long turnaround times and off-site testing.
• Rapid, on-site molecular diagnostics are needed for critical clinical scenarios.

Purpose of the Study:

• To develop a rapid, multiplex molecular method for identifying clinically relevant blood group alleles.
• To create an easy-to-use, on-site assay for blood group genotyping.
• To address the limitations of current molecular typing methods in emergency transfusion medicine.

Main Methods:

• Development of a dry-reagent, allele-specific lateral flow biosensor.
• Optimization of multiplex linear-after-the-exponential (LATE)-PCR and lateral flow parameters.
• Assay design to eliminate the need for DNA extraction, enabling direct use with blood samples.

Main Results:

• The novel assay identifies seven alleles across the Kidd, Duffy, and MNS blood group systems.
• The entire process, from sample receipt to result, takes approximately one hour.
• Achieved 100% concordance between deduced genotypes and standard serological phenotypes in 108 blood donors.
• Demonstrated ease of use, suitable for non-skilled healthcare professionals.

Conclusions:

• The developed rapid multiplex molecular assay provides an accurate and efficient alternative for blood group genotyping.
• The assay's speed, simplicity, and on-site capability enhance its utility in emergency transfusion settings.
• This platform has potential for broader applications in immunohematology and infectious disease diagnostics using single nucleotide polymorphism (SNP) panels.