Korean J Blood Transfus.  2022 Dec;33(3):154-160. 10.17945/kjbt.2022.33.3.1 5 4.

Kidd (SLC14A1 rs1058396) Genotyping Performed by PCR-Restriction Fragment Length Polymorphism Using a General-Purpose Agarose Gel and Lithium Borate Buffer

• Park G

• KMID: 2537221
• DOI: http://doi.org/10.17945/kjbt.2022.33.3.1 5 4

Abstract

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.

Keyword

Kidd genotyping; SLC14A1 rs1058396; RFLP; Lithium borate buffer; Agarose