High-throughput Kidd blood group genotyping based on molecular inversion probe and next generation sequencing technology
Objective:To develop a molecular inversion probe(MIP)capture for targeted next generation se-quencing(NGS)based to allow simultaneous identification of the Kidd blood group in multiple patients in a single run.Methods:After DNA extraction,MIP probes were used to capture the targets.Primers with sample bar-codes and Illumina adaptors were used to amplified the products by polymerase chain reaction(PCR).Samples were sequenced using an Illumina MiSeq.Three samples from blood donors previously test by PCR-SSP assay were used for comparison testing to determine if the established method could distinguish J KA and J KB 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 were the same as those of the PCR-SSP method,indicating that the new method 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 were 0.39 and 0.61 in the 100 individuals,and the antigen frequency was consistent with its distribution in the Eastern population.Conclu-sion:The MIP-based NGS test may be 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 in-corporation of new capture probes.
kidd blood groupnext generation sequencingmolecular inversion probeblood group genotyping
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