범용 아가로스 겔과 리튬붕산염버퍼를 이용한 PCR-제한효소절편길이다형성법으로 실시한 Kidd (SLC14A1 rs1058396) 유전형 검사 (Kidd (SLC14A1 rs1058396) Genotyping Performed by PCR-Restriction Fragment Length Polymorphism Using a General-Purpose Agarose Gel and Lithium Borate Buffer)

서지정보

· 발행기관 : 대한수혈학회
· 수록지 정보 : 대한수혈학회지 / 33권 / 3호 / 154 ~ 160페이지
· 저자명 : 박건

초록

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. (Korean J Blood Transfus 2022;33:154-160)

영어초록

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. (Korean J Blood Transfus 2022;33:154-160)

참고자료

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