High-throughput Kidd Blood Group Genotyping Based On Molecular Inversion Probe

Objective Serology is the traditional method for Kidd blood group typing.However,this technique has some limitations and cannot respond to the growing demand for Kidd antigens typing.This study aimed to develop a molecular inversion probe(MIP)capture for targeted Next generation sequencing(NGS)based to allow simultaneous identification of the Kidd blood group in multiple patients in a single run.Method After DNA extraction,MIP probes are used to capture the targets.Primers with sample barcodes and Illumina adaptors was used to amplified the products by polymerase chain reaction(PCR).Samples were sequenced using an Illumina Next Seq.Three samples from blood donors previously test by PCR-SSP assay were used for comparison testing to determine if the established method could distinguish JKa and JKb alleles effectively.Then,MIP based NGS results were compared to serologic for 100 samples.Results The results of the MIP-based NGS blood typing method are the same as those of the PCR-SSP method,indicating that the new method can could distinguish JKa and JKb alleles effectively.MIP based NGS red blood cell antigen typing was 100%concordant with serologic and SNP based typing.The frequencies of JKa and JKb alleles were 0.39 and 0.61 in the 100 individuals,and the antigen frequency was consistent with its distribution in the eastern population.Conclusion The MIP-based NGS test is a viable blood typing strategy that allows for large-scale Kidd blood group testing.Furthermore,extending this overall MIP approach to new blood group systems will only require the incorporation of new capture probes.