A direct blood polymerase chain reaction approach for the determination of GP.Mur (Mi.III) and other Hil+ Miltenberger glycophorin variants

Authors

  • Kate Hsu,

    Corresponding author
    1. From the Mackay Memorial Hospital Transfusion Medicine Laboratory, Tamsui; and the Department of Hematology-Oncology, Mackay Memorial Hospital, and the Mackay Memorial Hospital Blood Bank, Taipei, Taiwan.
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    • These authors contributed equally to this work.

  • Yen-Chun Lin,

    1. From the Mackay Memorial Hospital Transfusion Medicine Laboratory, Tamsui; and the Department of Hematology-Oncology, Mackay Memorial Hospital, and the Mackay Memorial Hospital Blood Bank, Taipei, Taiwan.
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    • These authors contributed equally to this work.

  • Yu-Cheng Chang,

    1. From the Mackay Memorial Hospital Transfusion Medicine Laboratory, Tamsui; and the Department of Hematology-Oncology, Mackay Memorial Hospital, and the Mackay Memorial Hospital Blood Bank, Taipei, Taiwan.
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  • Yung-Syu Chan,

    1. From the Mackay Memorial Hospital Transfusion Medicine Laboratory, Tamsui; and the Department of Hematology-Oncology, Mackay Memorial Hospital, and the Mackay Memorial Hospital Blood Bank, Taipei, Taiwan.
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  • Hsueh-Ping Chao,

    1. From the Mackay Memorial Hospital Transfusion Medicine Laboratory, Tamsui; and the Department of Hematology-Oncology, Mackay Memorial Hospital, and the Mackay Memorial Hospital Blood Bank, Taipei, Taiwan.
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  • Ting-Ying Lee,

    1. From the Mackay Memorial Hospital Transfusion Medicine Laboratory, Tamsui; and the Department of Hematology-Oncology, Mackay Memorial Hospital, and the Mackay Memorial Hospital Blood Bank, Taipei, Taiwan.
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  • Marie Lin

    1. From the Mackay Memorial Hospital Transfusion Medicine Laboratory, Tamsui; and the Department of Hematology-Oncology, Mackay Memorial Hospital, and the Mackay Memorial Hospital Blood Bank, Taipei, Taiwan.
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  • This work was supported by grants from Taiwan National Science Council (NSC 99-2320-B195-001-MY3), National Health Research Institute (NHRI-EX101-10122SI), and Mackay Memorial Hospital to KH.

Kate Hsu, Mackay Memorial Hospital Transfusion Medicine Laboratory, 45 Min-Sheng Road, Research Building 616, Tamsui, Taiwan 251; e-mail: khsu1@ms1.mmh.org.tw.

Abstract

BACKGROUND: GP.Mur (Mi.III) is a glycophorin B-A-B hybrid sialoglycoprotein expressing several potent immunogens, including Mia, Mur, and Hil. GP.Mur is considered one of the most important red blood cell (RBC) phenotypes in blood banking in Southeast Asia. However, there are no antibodies commercially available for the screening of GP.Mur RBCs.

STUDY DESIGN AND METHODS: To develop a direct blood polymerase chain reaction (PCR) approach for the screening of GP.Mur cells, we first confirmed the genomic sequence differences among four GP.Mur and three Mi(a–) samples by sequencing their GYP.Mur and GYPB genes. With these data, we designed PCR primers that best discriminate GYPB and GYP.Mur. Our primer design also allows the detection of other Hil+ glycophorin variants. We also constructed two plasmids—pGBi2i3 and pMiIIIi2i3—which serve as the negative and positive control DNA, respectively, for the PCR procedure. Additionally, we designed a control PCR to be run side by side with the typing PCR.

RESULTS: Because of the high specificity of our primers, we found it unnecessary to extract DNA from blood samples for PCR. We have tested this PCR method on 379 fresh and frozen blood samples. The results were further validated by serology and DNA sequencing and were shown to be completely accurate in our hand. We also found that the rapid genotyping method—high-resolution melting—can be a timesaving alternative for DNA sequencing.

CONCLUSION: This direct blood PCR approach for determination of GP.Mur and related Hil+ phenotypes is reliable and economical and is expected to be useful for blood banking in Southeast Asia.

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