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      MNSs Blood Group Glycophorin Variants in Taiwan: A Genotype-Serotype Correlation Study of ‘Mi a’ and St a with Report of Two New Alleles for St a

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          Abstract

          Background

          Glycophorin variants of the MNSs blood group are important in Taiwan. For more than 20 years, screening for the most frequent irregular antibody, anti-‘Mi a’, has been conducted by using ‘Mi a’(+) RBCs, with a significant success. However, the sensitivity and the specificity of this screening strategy have never been validated, and the true incidences of different glycophorin variants in Taiwan have been in controversy. Also, the significance of another less frequent and usually separately reported variant, St a, has never been evaluated.

          Methodology/Principal Findings

          We ran a population-based screening (from unselected patients in our hospital) for MNSs blood group glycophorin variants by PCR-sequencing method. GP.Mur (Mil.III) was confirmed by sequence from 57 out of 1027 samples (5.6%), and there was no other Miltenberger subtype glycophorin variant found. Glycophorin variant St a was found from 35 out of 1027 samples (3.4%). In contrast to anti-‘Mi a’, which is the most frequently identified irregular antibody in Taiwan, the prevalence of anti-St a was only 0.13% as determined by serologic method. In addition, two new alleles for St a were found and reported.

          Conclusion/Significance

          We confirm the long-standing assumption that GP.Mur is the only prevalent Miltenberger subtype in Taiwan. The current anti-‘Mi a’ screening method used in Taiwan, although neither sensitive nor specific, is still a suitable practice. Although St a antigen has a high prevalence in Taiwan, routine screening for anti-St a is not warranted based on current evidence.

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          Most cited references9

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

          GP.Mur (Mi.III) is a glycophorin B-A-B hybrid sialoglycoprotein expressing several potent immunogens, including Mi(a), 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. 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. 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. 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. © 2012 American Association of Blood Banks.
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            A survey of the incidence of Miltenberger antibodies among Hong Kong Chinese blood donors.

            The ready availability of red cells of the Miltenberger (Mi) class III phenotype (6.28%) prompted the study of Mi antibodies among Chinese blood donors in Hong Kong, 98 percent of whom are descended from inhabitants of Guangdong Province in southern China. Red cells of the Mi class III phenotype were used to conduct a survey of the frequency of Miltenberger antibodies in 56,161 random Chinese blood donors, over a period of 12 months, using a microplate technique. Sera from 32 donors (0.057%) were found to contain Mi antibodies: sera from 22 contained anti-Mur + Hut; sera from 4 contained anti-Vw + Mur + Hut; sera from 4 had monospecific anti-Mur; and sera from 2 had monospecific anti-Hil. The immunoglobulin isotypes of 24 sera were mixtures of IgM and IgG, 4 were pure IgM, and 4 were pure IgG. The majority of Mi antibodies detected were naturally occurring. This survey proved useful for mass screening of random donors for the procurement of valuable Mi antisera.
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              Detection of MNS hybrid molecules in the Thai population using PCR-SSP technique.

              We developed a polymerase chain reaction-sequence-specific primer (PCR-SSP) technique to screen for hybrid molecules in the MNS blood group in the Thai population using two sets of newly designed primers specific for four GYP(B-A-B) hybrids, GP.Mur, GP.Hop, GP.Bun and GP.HF, and two GYP(A-B-A) hybrids, GP.Vw and GP.Hut. One thousand and forty-one blood samples were tested with human anti-Mi(a) by conventional tube technique, and 598 samples of these were tested by the PCR-SSP technique. Ninety-four samples (9.03%) were strongly positive with human antisera by conventional tube technique. For PCR-SSP test results, the GP.Hut, GP.Mur, GP.Hop, GP.Bun and GP.HF genotypes were amplified with the first set of primers, whereas GP.Vw genotype was amplified with a second set of primers. The GYP(A-B) hybrids (GP. Hil and GP.JL), GYP(A-B-A) hybrids (GP.Nob, GP.Joh and GP.Dane), GYPA, GYPB and GYPE were not amplified by either set of primers. Results of testing 94 Mi(a+) and 504 Mi(a-) by conventional tube technique and PCR-SSP were concordant. This study shows that analysis by PCR-SSP is simple and convenient; therefore, it can be used as an alternative to conventional tube technique for mass screening for MNS hybrids, especially when specific antisera are not available.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                23 May 2014
                : 9
                : 5
                : e98166
                Affiliations
                [1 ]Department of Anatomic Pathology, Chang Gung Memorial Hospital, Taoyuan County, Taiwan
                [2 ]Department of Laboratory Medicine, Chang Gung Memorial Hospital, Taoyuan County, Taiwan
                [3 ]Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan County, Taiwan
                [4 ]Department of Pathology, School of Medicine, Chang Gung University, Taoyuan County, Taiwan
                German Red Cross Blood Service Frankfurt, Germany
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: TDC DPC WTW CFS. Performed the experiments: TDC WTW. Analyzed the data: TDC CFS. Contributed reagents/materials/analysis tools: DPC WTW CFS. Wrote the paper: TDC CFS.

                Article
                PONE-D-14-03425
                10.1371/journal.pone.0098166
                4032321
                24858913
                cdfbd218-2d38-477e-a180-210215b1c9f7
                Copyright @ 2014

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 26 January 2014
                : 29 April 2014
                Page count
                Pages: 5
                Funding
                This work was supported by the Grant CMRPG300091-93 from the Chang Gung Memorial Hospital, Taiwan. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Medicine and Health Sciences
                Diagnostic Medicine
                Clinical Laboratory Sciences
                Transfusion Medicine
                Hematology

                Uncategorized
                Uncategorized

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