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KMID : 0387720220330030154
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Korean Journal of Blood Transfusion
2022 Volume.33 No. 3 p.154 ~ p.160
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Kidd (SLC14A1 rs1058396) Genotyping Performed by PCR-Restriction Fragment Length Polymorphism Using a General-Purpose Agarose Gel and Lithium Borate Buffer
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Park Geon
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Abstract
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Background:Kidd (SLC14A1 rs1058396) genotyping is an important test to prevent delayed hemolytic transfusion reactions and hemolytic disease of the fetus or newborn. The PCR-restriction fragment length polymorphism (RFLP) technique using the MnlI restriction enzyme is not used widely because of the need for a polyacrylamide gel. PCR-RFLP was performed by electrophoresis with general-purpose agarose gel, and lithium borate buffer (LBB) was developed as a replacement for polyacrylamide gel.
Methods:Seventy-two venous blood samples were collected randomly and used in this study. A 3% agarose gel containing 1 μg/mL of ethidium bromide and 1 mM LBB, and a high-voltage (300 V) were used to separate the short-length restriction fragments (72 bp, 51 bp, 36 bp, and 21 bp). The PCR-RFLP results were confirmed by PCR-direct sequencing.
Results:Target restriction fragments could be easily discriminated. The results obtained with the PCR-RFLP were completely in agreement with the results of PCR-direct sequencing.
Conclusion:The PCR-RFLP using a general-purpose agarose gel and LBB is an accurate and reliable assay for Kidd genotyping.
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KEYWORD
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Kidd genotyping, SLC14A1 rs1058396, RFLP, Lithium borate buffer, Agarose
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