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(PDF) Genotyping analysis of MNS blood group GP(B‐A‐B) hybrid glycophorins in the Chinese Southern Han population using a high‐resolution melting assay
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"https://www.academia.edu/login?post_login_redirect_url=https%3A%2F%2Fwww.academia.edu%2F115701563%2FGenotyping_analysis_of_MNS_blood_group_GP_B_A_B_hybrid_glycophorins_in_the_Chinese_Southern_Han_population_using_a_high_resolution_melting_assay%3Fshow_translation%3Dtrue"; window.loswp.previewableAttachments = [{"id":112034700,"identifier":"Attachment_112034700","shouldShowBulkDownload":false}]; window.loswp.shouldDetectTimezone = true; window.loswp.shouldShowBulkDownload = true; window.loswp.showSignupCaptcha = false window.loswp.willEdgeCache = false; window.loswp.work = {"work":{"id":115701563,"created_at":"2024-03-02T21:57:10.034-08:00","from_world_paper_id":250793662,"updated_at":"2024-11-24T14:01:56.873-08:00","_data":{"publisher":"Wiley-Blackwell","grobid_abstract":"BACKGROUND: MNS hybrid GP(B-A-B) glycophorins are more commonly found in Southeast Asians and alloantibodies to antigens they carry are clinically significant. Detection of hybrid glycophorins by serologic techniques is limited due to lack of commercial reagents. In this study, a genotyping method for GP(B-A-B) hybrid glycophorins based on high-resolution melting (HRM) analysis was applied for genotyping analysis in the Chinese Southern Han population. STUDY DESIGN AND METHODS: DNA samples from 3104 Chinese Southern Han blood donors were collected. GYP(B-A-B) genotypes were analyzed by HRM assay. Parts of samples (n 5 106) were also tested by multiplex ligation-dependent probe amplification (MLPA) assay. Direct sequencing was conducted in samples with variant melting curve profiles. RESULTS: A total of five GYP(B-A-B) genotypes (201/ 3104, 6.5%) were identified, which were GYP*Mur heterozygote (n 5 194), GYP*Mur homozygote (n 5 3), GYP*Bun heterozygote (n 5 2), GYP*HF heterozygote (n 5 1), and a novel GYP(B-A-B) hybrid allele (n 5 1). Genotyping results for GYP*Mur and wild-type GYPB samples obtained by HRM were consistent with MLPA, while GYP*Bun and GYP*HF heterozygote identified by HRM could only be identified to have one copy of 5 0 inactive splice site of GYPB Pseudoexon 3 by MLPA. In addition, 10 single-nucleotide polymorphisms (SNPs) including four known and six novel SNPs were identified in 31 samples. One sample was identified carrying both GYP*Mur and GYP*Sch alleles. CONCLUSION: The HRM assay could distinguish the GYP(B-A-B) hybrid alleles successfully. Polymorphisms identified within the GYPB gene should be taken into consideration when developing GYP(B-A-B) genotyping kits for the Chinese population.","publication_date":"2018,6,13","publication_name":"Transfusion","grobid_abstract_attachment_id":"112034700"},"document_type":"paper","pre_hit_view_count_baseline":null,"quality":"high","language":"en","title":"Genotyping analysis of MNS blood group GP(B‐A‐B) hybrid glycophorins in the Chinese Southern Han population using a high‐resolution melting assay","broadcastable":false,"draft":null,"has_indexable_attachment":true,"indexable":true}}["work"]; window.loswp.workCoauthors = [40694093]; window.loswp.locale = "en"; window.loswp.countryCode = "SG"; window.loswp.cwvAbTestBucket = ""; window.loswp.designVariant = "ds_vanilla"; window.loswp.fullPageMobileSutdModalVariant = "full_page_mobile_sutd_modal"; window.loswp.useOptimizedScribd4genScript = false; window.loginModal = {}; window.loginModal.appleClientId = 'edu.academia.applesignon'; window.userInChina = "false";</script><script defer="" src="https://accounts.google.com/gsi/client"></script><div class="ds-loswp-container"><div class="ds-work-card--grid-container"><div class="ds-work-card--container js-loswp-work-card"><div class="ds-work-card--cover"><div class="ds-work-cover--wrapper"><div class="ds-work-cover--container"><button class="ds-work-cover--clickable js-swp-download-button" data-signup-modal="{"location":"swp-splash-paper-cover","attachmentId":112034700,"attachmentType":"pdf"}"><img alt="First page of “Genotyping analysis of MNS blood group GP(B‐A‐B) hybrid glycophorins in the Chinese Southern Han population using a high‐resolution melting assay”" class="ds-work-cover--cover-thumbnail" src="https://0.academia-photos.com/attachment_thumbnails/112034700/mini_magick20240303-1-tna7cc.png?1709445533" /><img alt="PDF Icon" class="ds-work-cover--file-icon" src="//a.academia-assets.com/images/single_work_splash/adobe_icon.svg" /><div class="ds-work-cover--hover-container"><span class="material-symbols-outlined" style="font-size: 20px" translate="no">download</span><p>Download Free PDF</p></div><div class="ds-work-cover--ribbon-container">Download Free PDF</div><div class="ds-work-cover--ribbon-triangle"></div></button></div></div></div><div class="ds-work-card--work-information"><h1 class="ds-work-card--work-title">Genotyping analysis of MNS blood group GP(B‐A‐B) hybrid glycophorins in the Chinese Southern Han population using a high‐resolution melting assay</h1><div class="ds-work-card--work-authors ds-work-card--detail"><a class="ds-work-card--author js-wsj-grid-card-author ds2-5-body-md ds2-5-body-link" data-author-id="40694093" href="https://independent.academia.edu/FlowerRobert"><img alt="Profile image of Robert Flower" class="ds-work-card--author-avatar" src="//a.academia-assets.com/images/s65_no_pic.png" />Robert Flower</a></div><div class="ds-work-card--detail"><p class="ds-work-card--detail ds2-5-body-sm">2018, Transfusion</p><div class="ds-work-card--work-metadata"><div class="ds-work-card--work-metadata__stat"><span class="material-symbols-outlined" style="font-size: 20px" translate="no">visibility</span><p class="ds2-5-body-sm" id="work-metadata-view-count">…</p></div><div class="ds-work-card--work-metadata__stat"><span class="material-symbols-outlined" style="font-size: 20px" translate="no">description</span><p class="ds2-5-body-sm">9 pages</p></div><div class="ds-work-card--work-metadata__stat"><span class="material-symbols-outlined" style="font-size: 20px" translate="no">link</span><p class="ds2-5-body-sm">1 file</p></div></div><script>(async () => { const workId = 115701563; 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if (!viewCountBody) { throw new Error('Failed to find work views element'); } viewCountBody.textContent = `${commaizedViewCount} views`; } catch (error) { // Remove the whole views element if there was some issue parsing. document.getElementById('work-metadata-view-count')?.parentNode?.remove(); throw new Error(`Failed to parse view count: ${viewCount}`, error); } }; // If the DOM is still loading, wait for it to be ready before updating the view count. if (document.readyState === "loading") { document.addEventListener('DOMContentLoaded', () => { updateViewCount(viewCount); }); // Otherwise, just update it immediately. } else { updateViewCount(viewCount); } })();</script></div><p class="ds-work-card--work-abstract ds-work-card--detail ds2-5-body-md">BACKGROUND: MNS hybrid GP(B-A-B) glycophorins are more commonly found in Southeast Asians and alloantibodies to antigens they carry are clinically significant. Detection of hybrid glycophorins by serologic techniques is limited due to lack of commercial reagents. In this study, a genotyping method for GP(B-A-B) hybrid glycophorins based on high-resolution melting (HRM) analysis was applied for genotyping analysis in the Chinese Southern Han population. STUDY DESIGN AND METHODS: DNA samples from 3104 Chinese Southern Han blood donors were collected. GYP(B-A-B) genotypes were analyzed by HRM assay. Parts of samples (n 5 106) were also tested by multiplex ligation-dependent probe amplification (MLPA) assay. Direct sequencing was conducted in samples with variant melting curve profiles. RESULTS: A total of five GYP(B-A-B) genotypes (201/ 3104, 6.5%) were identified, which were GYP*Mur heterozygote (n 5 194), GYP*Mur homozygote (n 5 3), GYP*Bun heterozygote (n 5 2), GYP*HF heterozygote (n 5 1), and a novel GYP(B-A-B) hybrid allele (n 5 1). Genotyping results for GYP*Mur and wild-type GYPB samples obtained by HRM were consistent with MLPA, while GYP*Bun and GYP*HF heterozygote identified by HRM could only be identified to have one copy of 5 0 inactive splice site of GYPB Pseudoexon 3 by MLPA. In addition, 10 single-nucleotide polymorphisms (SNPs) including four known and six novel SNPs were identified in 31 samples. One sample was identified carrying both GYP*Mur and GYP*Sch alleles. CONCLUSION: The HRM assay could distinguish the GYP(B-A-B) hybrid alleles successfully. Polymorphisms identified within the GYPB gene should be taken into consideration when developing GYP(B-A-B) genotyping kits for the Chinese population.</p><div class="ds-work-card--button-container"><button class="ds2-5-button js-swp-download-button" data-signup-modal="{"location":"continue-reading-button--work-card","attachmentId":112034700,"attachmentType":"pdf","workUrl":"https://www.academia.edu/115701563/Genotyping_analysis_of_MNS_blood_group_GP_B_A_B_hybrid_glycophorins_in_the_Chinese_Southern_Han_population_using_a_high_resolution_melting_assay"}">See full PDF</button><button class="ds2-5-button ds2-5-button--secondary js-swp-download-button" data-signup-modal="{"location":"download-pdf-button--work-card","attachmentId":112034700,"attachmentType":"pdf","workUrl":"https://www.academia.edu/115701563/Genotyping_analysis_of_MNS_blood_group_GP_B_A_B_hybrid_glycophorins_in_the_Chinese_Southern_Han_population_using_a_high_resolution_melting_assay"}"><span class="material-symbols-outlined" style="font-size: 20px" translate="no">download</span>Download PDF</button></div><div class="ds-signup-banner-trigger-container"><div class="ds-signup-banner-trigger ds-signup-banner-trigger-control"></div></div><div class="ds-signup-banner ds-signup-banner-control"><div id="ds-signup-banner-close-button"><button class="ds2-5-button ds2-5-button--secondary ds2-5-button--inverse"><span class="material-symbols-outlined" style="font-size: 20px" translate="no">close</span></button></div><div class="ds-signup-banner-ctas"><img src="//a.academia-assets.com/images/academia-logo-capital-white.svg" /><h4 class="ds2-5-heading-serif-sm">Sign up for access to the world's latest research</h4><button class="ds2-5-button ds2-5-button--inverse ds2-5-button--full-width js-swp-download-button" data-signup-modal="{"location":"signup-banner"}">Sign up for free<span class="material-symbols-outlined" style="font-size: 20px" translate="no">arrow_forward</span></button></div><div class="ds-signup-banner-divider"></div><div class="ds-signup-banner-reasons"><div class="ds-signup-banner-reasons-item"><span class="material-symbols-outlined" style="font-size: 24px" translate="no">check</span><span>Get notified about relevant papers</span></div><div class="ds-signup-banner-reasons-item"><span class="material-symbols-outlined" style="font-size: 24px" translate="no">check</span><span>Save papers to use in your research</span></div><div class="ds-signup-banner-reasons-item"><span class="material-symbols-outlined" style="font-size: 24px" translate="no">check</span><span>Join the discussion with peers</span></div><div class="ds-signup-banner-reasons-item"><span class="material-symbols-outlined" style="font-size: 24px" translate="no">check</span><span>Track your impact</span></div></div></div><script>(() => { // Set up signup banner show/hide behavior: // 1. 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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.</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"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","attachmentId":44389369,"attachmentType":"pdf","work_url":"https://www.academia.edu/24006109/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","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/24006109/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"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="1" data-entity-id="60900886" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/60900886/A_minisatellite_and_a_microsatellite_polymorphism_within_1_5_kb_at_the_human_muscle_glycogen_phosphorylase_PYGM_locus_can_be_amplified_by_PCR_and_have_combined_informativeness_of_PIC_0_95">A minisatellite and a microsatellite polymorphism within 1.5 kb at the human muscle glycogen phosphorylase (PYGM) locus can be amplified by PCR and have combined informativeness of PIC 0.95</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="38110719" href="https://independent.academia.edu/HDoniskeller">H. Donis-keller</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Genomics, 1992</p><p class="ds-related-work--abstract ds2-5-body-sm">We sequenced a genomic clone (pMCMPl), previously reported to detect a VNTR polymorphism at the PYGM locus, and found a dinucleotide repeat segment (CA),,(GA),, and a complex (AT)-repeat-rich segment containing 63 repeats spanning 160 bp. Resolution of PCR-amplified genomic DNA from the (CA)(GA) repeat region on DNA sequencing gels revealed a highly informative polymorphism with alleles differing by Z-bp intervals and ranging in size from 156 to 190 bp. Among three racial groups, a total of 18 alleles were observed. Fourteen alleles were observed in Caucasians (PIC O&9), 12 alleles in American Blacks (PIC 0.89), and 9 alleles in Pima Indians (PIC 0.73). PCR amplification of the (AT) repeat region and resolution of the products on DNA sequencing gels revealed a complex variable length polymorphism with alleles distributed in size from 367 to 970 bp. Twenty-eight alleles were found in American Blacks (PIC 0.94), 6 alleles in Pima Indians (PIC 0.70), and 11 alleles in Caucasians (PIC 0.71). Comparison of the previously described VNTR RFLP alleles visualized by Southern hybridization to the PCR products described in this report demonstrated that the polymorphism detected in both assays was identical. However, a larger number of alleles could be detected from the PCR-amplified products. Combined informativeness, PIC 0.95, for the two polymorphisms was determined from haplotype analysis of 100 Caucasian chromosomes. Therefore, for genotyping purposes, informativeness is maximized from using both polymorphisms. The sequence of the PYGM 2.1-kb cloned insert has been deposited with the GenBank Data Libraries (Accession No. M77201).</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"A minisatellite and a microsatellite polymorphism within 1.5 kb at the human muscle glycogen phosphorylase (PYGM) locus can be amplified by PCR and have combined informativeness of PIC 0.95","attachmentId":74143261,"attachmentType":"pdf","work_url":"https://www.academia.edu/60900886/A_minisatellite_and_a_microsatellite_polymorphism_within_1_5_kb_at_the_human_muscle_glycogen_phosphorylase_PYGM_locus_can_be_amplified_by_PCR_and_have_combined_informativeness_of_PIC_0_95","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/60900886/A_minisatellite_and_a_microsatellite_polymorphism_within_1_5_kb_at_the_human_muscle_glycogen_phosphorylase_PYGM_locus_can_be_amplified_by_PCR_and_have_combined_informativeness_of_PIC_0_95"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="2" data-entity-id="110536876" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/110536876/Validation_of_the_multiplex_ligation_dependent_probe_amplification_assay_and_its_application_on_the_distribution_study_of_the_major_alleles_of_17_blood_group_systems_in_scp_C_scp_hinese_donors_from_scp_G_scp_uangzhou">Validation of the multiplex ligation‐dependent probe amplification assay and its application on the distribution study of the major alleles of 17 blood group systems in <scp>C</scp> hinese donors from <scp>G</scp> uangzhou</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="86894093" href="https://independent.academia.edu/LonnekeHaerWigman">Lonneke Haer-Wigman</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Transfusion, 2016</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Validation of the multiplex ligation‐dependent probe amplification assay and its application on the distribution study of the major alleles of 17 blood group systems in \u003cscp\u003eC\u003c/scp\u003e hinese donors from \u003cscp\u003eG\u003c/scp\u003e uangzhou","attachmentId":108324462,"attachmentType":"pdf","work_url":"https://www.academia.edu/110536876/Validation_of_the_multiplex_ligation_dependent_probe_amplification_assay_and_its_application_on_the_distribution_study_of_the_major_alleles_of_17_blood_group_systems_in_scp_C_scp_hinese_donors_from_scp_G_scp_uangzhou","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/110536876/Validation_of_the_multiplex_ligation_dependent_probe_amplification_assay_and_its_application_on_the_distribution_study_of_the_major_alleles_of_17_blood_group_systems_in_scp_C_scp_hinese_donors_from_scp_G_scp_uangzhou"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="3" data-entity-id="52951221" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/52951221/Characterization_of_GYP_Mur_and_novel_GYP_Bun_like_hybrids_in_Thai_blood_donors_reveals_a_qualitatively_altered_s_antigen">Characterization of GYP*Mur and novel GYP*Bun -like hybrids in Thai blood donors reveals a qualitatively altered s antigen</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="32964155" href="https://quotientbioresearch.academia.edu/JanineRobb">Janine Robb</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Vox Sanguinis</p><p class="ds-related-work--abstract ds2-5-body-sm">Background and objectives The Mi(a+) GP(B-A-B) hybrid phenotypes occur with a prevalence of 2%-23% across Southeast Asia. While the s antigen is alleged to be altered, no evidence for specific variants is known. Screening using a monoclonal IgM anti-s mistyped six S-s+ RBC units as S-s-. Further, alloanti-s was identified in an S+s+ patient. Our objective was to investigate the s antigen further. Materials and methods DNA from 63 Thai blood donor samples PCR-positive for a GYP(B-A-B) hybrid was sequenced with primers spanning GYPB exons 3-4. Flow cytometry was used for semiquantitative analysis of s expression and correlated with the glycophorin genotype. Results DNA sequencing showed that GYP*Mur was carried by 56/63 samples (88Á9%) of which 5/56 lacked normal GYPB: three of these were GYP*Mur homozygotes, one was a compound heterozygote carrying GYP*Mur and a GYP*-Bun-like allele (designated GYP*Thai), and the fifth sample carried GYP*Mur and another GYP*Bun-like allele. Seven samples (7/63) were GYP*Thai heterozygotes. IgM monoclonal anti-s (P3BER) did not react with the s antigen carried by GP.Mur or GP.Bun, whereas two IgG anti-s showed enhanced reactivity. Conclusions We confirmed that GYP*Mur is the most frequent variant in Thai blood donors and also identified GYP*Thai with a frequency of 1Á1%. We showed that s antigen on Mi(a+) GP(B-A-B) hybrids is qualitatively altered and should be considered when selecting reagents for phenotyping where such hybrids are prevalent, endemically and in blood centres worldwide.</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Characterization of GYP*Mur and novel GYP*Bun -like hybrids in Thai blood donors reveals a qualitatively altered s antigen","attachmentId":69965440,"attachmentType":"pdf","work_url":"https://www.academia.edu/52951221/Characterization_of_GYP_Mur_and_novel_GYP_Bun_like_hybrids_in_Thai_blood_donors_reveals_a_qualitatively_altered_s_antigen","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/52951221/Characterization_of_GYP_Mur_and_novel_GYP_Bun_like_hybrids_in_Thai_blood_donors_reveals_a_qualitatively_altered_s_antigen"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="4" data-entity-id="90960520" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/90960520/Polymerase_chain_reaction_based_detection_of_MN_blood_group_specific_sequences_in_the_human_genome">Polymerase chain reaction-based detection of MN blood group-specific sequences in the human genome</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="140954821" href="https://independent.academia.edu/VarunBassessar">Varun Bassessar</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Transfusion, 1993</p><p class="ds-related-work--abstract ds2-5-body-sm">The MN blood group antigens have traditionally been detected by serotyping; however, development of a DNA-based method offers flexibility in the determination of this hi hly polymorphic system. Genotyping the MN blood grou antigens was performe! by polymerase chain reaction amplification of the speci& alleles (PASA) in the human genome. In separate paired reactions, M or N allele-specific oligonucleotide primers were amplified with a common distal primer. Only in the presence of the homologous template was a 781-base pair polymerase chain reaction amplification product visible after agarose gel electrophoresis and ethidium bromide staining. This method of genotyping could be performed using either 1 pg of extracted DNA or 0.5 pL of whole blood, and the results showed 100-percent correlation with those obtained by sero ping. PASA-based genotyping of MN blood group antigens, whjch studies and in forensic medicine. TRANSFUSION 1993;33:119-124. requires a smal Y amount of starting material, has application in linkage and population Abbrevlatlone: ASOP(8) = allele-speclflc ollgonucleotlde prlmer(8); bp = base pairs; QPA = glycophorln A; GPB = glycophorln B; GPC = glycophorln C; GPD = glycophorln D; OPE = glycophorln E; PASA = polymerase chaln reactlon ampllflcatlon of specific alleles; PCR = polymerase chaln reactlon.</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Polymerase chain reaction-based detection of MN blood group-specific sequences in the human genome","attachmentId":94380303,"attachmentType":"pdf","work_url":"https://www.academia.edu/90960520/Polymerase_chain_reaction_based_detection_of_MN_blood_group_specific_sequences_in_the_human_genome","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/90960520/Polymerase_chain_reaction_based_detection_of_MN_blood_group_specific_sequences_in_the_human_genome"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="5" data-entity-id="21381999" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/21381999/Single_PCR_Multiplex_SNaPshot_Reaction_for_Detection_of_Eleven_Blood_Group_Nucleotide_Polymorphisms">Single PCR Multiplex SNaPshot Reaction for Detection of Eleven Blood Group Nucleotide Polymorphisms</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="42457623" href="https://independent.academia.edu/JulieCristofaro">Julie Cristofaro</a></div><p class="ds-related-work--metadata ds2-5-body-xs">The Journal of Molecular Diagnostics, 2010</p><p class="ds-related-work--abstract ds2-5-body-sm">Hemagglutination-based assays have several clinical shortcomings. To overcome this difficulty , we have developed a multiplex-PCR SNaPshot assay adapted to the Southern French population , which includes individuals from sub-Saharan Africa and the Comoros archipelago. Single nucleotide polymorphisms (SNPs) associated with clinically relevant blood antigens as well as with null phenotypes were profiled (i.e. , K/k, ). A single multiplex-PCR reaction was used to amplify nine gene regions encompassing 11 SNPs. Identification was obtained by incorporation of the complementary dye-conjugated single base at the 3 end of each probe primer annealed proximal to the target SNP. After optimization, the SNaPshot assay was validated with 265 known allele or phenotype pairs. Results were found fully concordant with those of hemagglutination , allele-specific PCR , and/or sequencing. The assay was then evaluated on 227 blood samples in a clinical context. A total of 203 derived-phenotypes were generated , including 82 atypical phenotypes [i.e. , Fy(b؉ w ) (n ؍ 32); K ؉ (n ؍ 22); Co(b؉) (n ؍ 8); Yt(b؉) (n ؍ 18); S-s؉U؉ var (n ؍ 2) , 105 null phenotypes , i.e. , Fy(a-b-) (n ؍ 97); S-s-U-(n ؍ 6); S-s-U؉ var (n ؍ 2)] and sixteen Fypositive samples carried a FY*Fy allele. The findings show that this assay can provide a low-cost and fast genotyping tool well adapted to local ethnically mixed populations</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Single PCR Multiplex SNaPshot Reaction for Detection of Eleven Blood Group Nucleotide Polymorphisms","attachmentId":41843999,"attachmentType":"pdf","work_url":"https://www.academia.edu/21381999/Single_PCR_Multiplex_SNaPshot_Reaction_for_Detection_of_Eleven_Blood_Group_Nucleotide_Polymorphisms","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/21381999/Single_PCR_Multiplex_SNaPshot_Reaction_for_Detection_of_Eleven_Blood_Group_Nucleotide_Polymorphisms"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="6" data-entity-id="97979830" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/97979830/_i_Sfa_i_Nl_Polymorphism_Distinguishes_the_Alleles_of_the_Glycophorin_A_Locus_That_Determine_the_MN_Blood_Group"><i>Sfa</i>Nl Polymorphism Distinguishes the Alleles of the Glycophorin A Locus That Determine the MN Blood Group</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="19188274" href="https://independent.academia.edu/GrantStephen">Stephen Grant</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Acta Haematologica, 2009</p><p class="ds-related-work--abstract ds2-5-body-sm">The GPA gene is a single-copy autosomal gene coding for the major erythroid cell-specific sialoglycoprotein, gly cophorin A. This locus is part of a family of related genes, including GPB and GPE, that have been localized to 4q28-31 [1], The human GPA gene has been cloned and extensively characterized to reveal a mammalian gene of typical size and complexity consisting of seven coding exons and six introns extending over 40 kb. The gene is transcribed and processed to yield mRNA species of 1.0, 1.7, and 2.2 kb with identical coding regions specifying a cleavable hydrophobic leader peptide in addition to the 131 amino acids of the mature protein, differing only in the extent of the 3' untranslated region. GPA expression is detectable at the early erythroblast stage of erythropoiesis and is complete at the normoblast stage of differentiation. Bone marrow and peripheral blood reticulocytes as well as mature erythrocytes maintain a stable level of 0.5-1.0 x 106 GPA molecules on the cell surface. Two common GPA alleles are present in the human population, coding for proteins that differ at the first and fifth amino acids of the mature protein. These two allelic forms of GPA are expressed codominantly in heterozygous GPAM,N individ uals and serve to determine the MN blood group. These two alleles are approximately equally represented in the America population, with allele frequencies estimated to be 0.53:0.47 M:N for Caucasians and 0.49:0.51 for Afri can-Americans [2], The nucleotide sequences of the cloned GPAM and GPAN cDNAs have been determined [3-5], and, as shown in figure 1, the protein polymorphisms are attributable to KARGER</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"\u003ci\u003eSfa\u003c/i\u003eNl Polymorphism Distinguishes the Alleles of the Glycophorin A Locus That Determine the MN Blood Group","attachmentId":99455952,"attachmentType":"pdf","work_url":"https://www.academia.edu/97979830/_i_Sfa_i_Nl_Polymorphism_Distinguishes_the_Alleles_of_the_Glycophorin_A_Locus_That_Determine_the_MN_Blood_Group","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/97979830/_i_Sfa_i_Nl_Polymorphism_Distinguishes_the_Alleles_of_the_Glycophorin_A_Locus_That_Determine_the_MN_Blood_Group"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="7" data-entity-id="28499174" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/28499174/A_direct_blood_polymerase_chain_reaction_approach_for_the_determination_of_GP_Mur_Mi_III_and_other_Hil_Miltenberger_glycophorin_variants">A direct blood polymerase chain reaction approach for the determination of GP.Mur (Mi.III) and other Hil+ Miltenberger glycophorin variants</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="53678956" href="https://independent.academia.edu/YungSyuChan">Yung-Syu Chan</a><span>, </span><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="53447873" href="https://independent.academia.edu/KateHsu1">Kate Hsu</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Transfusion, 2013</p><p class="ds-related-work--abstract ds2-5-body-sm">BACKGROUND: 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. 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. ABBREVIATIONS: GYPA = glycophorin A; GYPB = glycophorin B; GYPE = glycophorin E; HRM = high-resolution melting.</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"A direct blood polymerase chain reaction approach for the determination of GP.Mur (Mi.III) and other Hil+ Miltenberger glycophorin variants","attachmentId":48847668,"attachmentType":"pdf","work_url":"https://www.academia.edu/28499174/A_direct_blood_polymerase_chain_reaction_approach_for_the_determination_of_GP_Mur_Mi_III_and_other_Hil_Miltenberger_glycophorin_variants","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/28499174/A_direct_blood_polymerase_chain_reaction_approach_for_the_determination_of_GP_Mur_Mi_III_and_other_Hil_Miltenberger_glycophorin_variants"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="8" data-entity-id="22872668" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/22872668/Analysis_of_multiple_single_nucleotide_polymorphisms_SNP_on_DNA_traces_from_plasma_and_dried_blood_samples">Analysis of multiple single nucleotide polymorphisms (SNP) on DNA traces from plasma and dried blood samples</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="32127392" href="https://independent.academia.edu/ChristinaVandenbrouckegrauls">Christina Vandenbroucke-Grauls</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Journal of Immunological Methods, 2007</p><p class="ds-related-work--abstract ds2-5-body-sm">Reliable analysis of single nucleotide polymorphisms (SNPs) in DNA derived from samples containing low numbers of cells or from suboptimal sources can be difficult. A new procedure to characterize multiple SNPs in traces of DNA from plasma and old dried blood samples was developed. Six SNPs in the Mannose Binding Lectin 2 (MBL2) gene were chosen as targets for analysis.</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Analysis of multiple single nucleotide polymorphisms (SNP) on DNA traces from plasma and dried blood samples","attachmentId":43415047,"attachmentType":"pdf","work_url":"https://www.academia.edu/22872668/Analysis_of_multiple_single_nucleotide_polymorphisms_SNP_on_DNA_traces_from_plasma_and_dried_blood_samples","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/22872668/Analysis_of_multiple_single_nucleotide_polymorphisms_SNP_on_DNA_traces_from_plasma_and_dried_blood_samples"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div><div class="ds-related-work--container js-wsj-grid-card" data-collection-position="9" data-entity-id="75901865" data-sort-order="default"><a class="ds-related-work--title js-wsj-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/75901865/High_throughput_genotyping_assays_for_identification_of_glycophorin_B_deletion_variants_in_population_studies">High-throughput genotyping assays for identification of glycophorin B deletion variants in population studies</a><div class="ds-related-work--metadata"><a class="js-wsj-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="13086988" href="https://independent.academia.edu/DAmuzu">Dominic Amuzu</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Experimental Biology and Medicine, 2020</p><p class="ds-related-work--abstract ds2-5-body-sm">Glycophorins are the most abundant sialoglycoproteins on the surface of human erythrocyte membranes. Genetic variation in glycophorin region of human chromosome 4 (containing GYPA, GYPB, and GYPE genes) is of interest because the gene products serve as receptors for pathogens of major public health interest, including Plasmodium sp., Babesia sp., Influenza virus, Vibrio cholerae El Tor Hemolysin, and Escherichia coli. A large structural rearrangement and hybrid glycophorin variant, known as Dantu, which was identified in East African populations, has been linked with a 40% reduction in risk for severe malaria. Apart from Dantu, other large structural variants exist, with the most common being deletion of the whole GYPB gene and its surrounding region, resulting in multiple different deletion forms. In West Africa particularly, these deletions are estimated to account for between 5 and 15% of the variation in different populations, mostly attributed to the forms known as DEL1 and DEL...</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"High-throughput genotyping assays for identification of glycophorin B deletion variants in population studies","attachmentId":83566589,"attachmentType":"pdf","work_url":"https://www.academia.edu/75901865/High_throughput_genotyping_assays_for_identification_of_glycophorin_B_deletion_variants_in_population_studies","alternativeTracking":true}"><span class="material-symbols-outlined" style="font-size: 18px" translate="no">download</span><span class="ds2-5-text-link__content">Download free PDF</span></button><a class="ds2-5-text-link ds2-5-text-link--inline js-wsj-grid-card-view-pdf" href="https://www.academia.edu/75901865/High_throughput_genotyping_assays_for_identification_of_glycophorin_B_deletion_variants_in_population_studies"><span class="ds2-5-text-link__content">View PDF</span><span class="material-symbols-outlined" style="font-size: 18px" translate="no">chevron_right</span></a></div></div></div></div><div class="ds-sticky-ctas--wrapper js-loswp-sticky-ctas hidden"><div class="ds-sticky-ctas--grid-container"><div class="ds-sticky-ctas--container"><button class="ds2-5-button js-swp-download-button" data-signup-modal="{"location":"continue-reading-button--sticky-ctas","attachmentId":112034700,"attachmentType":"pdf","workUrl":null}">See full PDF</button><button class="ds2-5-button ds2-5-button--secondary js-swp-download-button" data-signup-modal="{"location":"download-pdf-button--sticky-ctas","attachmentId":112034700,"attachmentType":"pdf","workUrl":null}"><span class="material-symbols-outlined" style="font-size: 20px" translate="no">download</span>Download PDF</button></div></div></div><div class="ds-below-fold--grid-container"><div class="ds-work--container js-loswp-embedded-document"><div class="attachment_preview" data-attachment="Attachment_112034700" style="display: none"><div class="js-scribd-document-container"><div class="scribd--document-loading js-scribd-document-loader" style="display: block;"><img alt="Loading..." src="//a.academia-assets.com/images/loaders/paper-load.gif" /><p>Loading Preview</p></div></div><div style="text-align: center;"><div class="scribd--no-preview-alert js-preview-unavailable"><p>Sorry, preview is currently unavailable. You can download the paper by clicking the button above.</p></div></div></div></div><div class="ds-sidebar--container js-work-sidebar"><div class="ds-related-content--container"><h2 class="ds-related-content--heading">Related papers</h2><div class="ds-related-work--container js-related-work-sidebar-card" data-collection-position="0" data-entity-id="123062619" data-sort-order="default"><a class="ds-related-work--title js-related-work-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/123062619/Multiplex_Universal_Genotyping_Using_a_Modified_ARMS_PCR_Protocol">Multiplex Universal Genotyping Using a Modified ARMS-PCR Protocol</a><div class="ds-related-work--metadata"><a class="js-related-work-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="42520929" href="https://independent.academia.edu/JVamvakopoulos">Joannis Vamvakopoulos</a></div><p class="ds-related-work--metadata ds2-5-body-xs">BioTechniques, 2002</p><div 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class="ds-related-work--container js-related-work-sidebar-card" data-collection-position="1" data-entity-id="118316580" data-sort-order="default"><a class="ds-related-work--title js-related-work-grid-card-title ds2-5-body-md ds2-5-body-link" href="https://www.academia.edu/118316580/Full_flexibility_genotyping_of_single_nucleotide_polymorphisms_by_the_GOOD_assay">Full flexibility genotyping of single nucleotide polymorphisms by the GOOD assay</a><div class="ds-related-work--metadata"><a class="js-related-work-grid-card-author ds2-5-body-sm ds2-5-body-link" data-author-id="199873380" href="https://independent.academia.edu/ChristinePlancon">Christine Plancon</a></div><p class="ds-related-work--metadata ds2-5-body-xs">Nucleic Acids Research, 2000</p><div class="ds-related-work--ctas"><button class="ds2-5-text-link ds2-5-text-link--inline js-swp-download-button" data-signup-modal="{"location":"wsj-grid-card-download-pdf-modal","work_title":"Full flexibility genotyping of single nucleotide 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