DOCSLIB.ORG

Targeted Exome Sequencing Defines Novel and Rare Variants in Complex Blood Group Serology Cases for a Red Blood Cell Reference Laboratory Setting

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Targeted Exome Sequencing Defines Novel and Rare Variants in Complex Blood Group Serology Cases for a Red Blood Cell Reference Laboratory Setting IMMUNOHEMATOLOGY Targeted exome sequencing defines novel and rare variants in complex blood group serology cases for a red blood cell reference laboratory setting Elizna M. Schoeman ,1 Eileen V. Roulis ,1 Yew-Wah Liew,2 Jacqueline R. Martin,2 Tanya Powley,2 Brett Wilson,2 Glenda M. Millard ,1 Eunike C. McGowan ,1 Genghis H. Lopez ,1 Helen O’Brien ,1 Jennifer A. Condon,3 Robert L. Flower ,1 and Catherine A. Hyland 1 he key role of a modern red blood cell (RBC) ref- BACKGROUND: We previously demonstrated that erence laboratory in transfusion medicine is to targeted exome sequencing accurately defined blood employ serology to solve complex problems. group genotypes for reference panel samples Often, the lack of appropriate RBCs, sera, or characterized by serology and single-nucleotide T polymorphism (SNP) genotyping. Here we investigate reagents makes confident resolution of a complex case the application of this approach to resolve problematic difficult. Resources can be costly and scarce, if available at serology and SNP-typing cases. all, and often require collaboration and generosity of col- 1 STUDY DESIGN AND METHODS: The TruSight One leagues from other international laboratories. In addition, sequencing panel and MiSeq platform was used for reference laboratories commonly employ single- sequencing. CLC Genomics Workbench software was nucleotide polymorphism (SNP) typing microarrays for used for data analysis of the blood group genes RBC genotyping investigations; however, these arrays are implicated in the serology and SNP-typing problem. not comprehensive in their coverage of blood groups or 2-5 Sequence variants were compared to public databases variants. When the combination of traditional serologic listing blood group alleles. The effect of predicted amino methods and typing by SNP microarray fails to resolve, acid changes on protein function for novel alleles was assessed using SIFT and PolyPhen-2. ABBREVIATIONS: ExAC 5 Exome Aggregation Consortium; RESULTS: Among 29 unresolved samples, sequencing MPS 5 massively parallel sequencing; RCRL 5 Red Cell defined SNPs in blood group genes consistent with Reference Laboratory; SNP(s) 5 single-nucleotide serologic observation: 22 samples exhibited SNPs polymorphism(s); SNV(s) 5 single-nucleotide variant(s); associated with varied but known blood group alleles and TSO 5 TruSight One. one sample exhibited a chimeric RH genotype. Three From 1Clinical Services and Research; and the 2Red Cell samples showed novel variants in the CROM, LAN, and Reference Laboratory, Australian Red Cross Blood Service, RH systems, respectively, predicting respective amino Kelvin Grove, Queensland, Australia; and the 3Red Cell acid changes with possible deleterious impact. Two Reference Laboratory, Australian Red Cross Blood Service, samples harbored rare variants in the RH and FY Melbourne, Victoria, Australia. systems, respectively, not previously associated with a Address reprint requests to: Catherine A. Hyland, PhD, blood group allele or phenotype. A final sample MSc, Clinical Services and Research, Australian Red Cross comprised a rare variant within the KLF1 transcription Blood Service, 44 Musk Avenue, Kelvin Grove, Queensland factor gene that may modulate DNA-binding activity. 4059, Australia; e-mail: [email protected]. CONCLUSION: Targeted exome sequencing resolved The Australian government funds the Australian Red Cross complex serology problems and defined both novel blood Blood Service for the provision of blood, blood products, and group alleles (CD55:c.203G>A, ABCB6:c.1118_1124del services to the Australian community. CGGATCG, ABCB6:c.1656-1G>A, and RHD:c.452G>A) Received for publication February 21, 2017; revision and rare variants on blood group alleles associated with al received August 21, 2017; and accepted September 5, 2017. tered phenotypes. This study illustrates the utility of exome doi:10.1111/trf.14393 sequencing, in conjunction with serology, as an alternative VC 2017 AABB approach to resolve complex cases. TRANSFUSION 2018;58;284–293 284 TRANSFUSION Volume 58, February 2018 EXOME SEQUENCING FOR COMPLEX BLOOD GROUPS further avenues of investigation are required. For some elution techniques were used to confirm presence of the reference laboratories the test algorithm at this stage will antigen. Cell separation of a dual population (Sample 7) include Sanger sequencing of targeted genes. was performed by agglutinating the D1 cells with anti-D As an alternative approach, massively parallel and separating from the unagglutinated cells using a 20% sequencing (MPS—also known as next-generation albumin gradient. sequencing) technologies are becoming more widely used Antibody identification was performed using stan- in blood banking and hematologic investigation, due to dard serologic techniques. Routinely, a commercial 11-cell the capability of identifying all variants in target blood panel (Seqirus) and autologous cells are tested by three group genes. A recent proof-of-principle study demon- tube methods: saline room temperature (248C), polyethyl- strated the capability of using whole genome sequencing ene glycol indirect antiglobulin test (IAT), and papain IAT. data for comprehensive antigen prediction of RBCs and Results from this routine testing directed further serologic platelets.6 A targeted exome sequencing approach has investigation with various enzymes or chemical modifica- been recognized for its utility in blood group genotyping tion of cells, rare phenotype cells, cord cells, antibody and has been predicted to facilitate investigation of novel adsorptions-elutions, and peptide inhibitions. blood group alleles.7,8 While studies have shown proof of principle for blood group genotyping by MPS, no studies SNP genotyping in the reference laboratory have investigated to date its clinical application to resolve Genomic DNA was extracted from fresh EDTA whole complex cases in a reference laboratory setting.6,8-12 blood samples using the DNA blood mini kit (QIAamp Previously, we validated the use of a commercially DSP, QIAGEN) for use with a robotic workstation for auto- available targeted exome sequencing panel for investiga- mated purification of DNA, RNA, or proteins (QIACube, tion of genetic variation within 28 protein-based blood QIAGEN) or manually as per the manufacturer’s instruc- group systems on a collection of well-characterized refer- tions if required. The DNA was tested using a high- ence panel samples.13 Our analyses confirmed the pheno- throughput molecular assay (BioArray BeadChip technol- types obtained using SNP microarray and serologic ogy, Immucor) for the simultaneous detection of a wide methods, as well as identifying a number of variants con- range of SNPs affecting RBC antigen expression, using a sidered “noninformative”—that is, they may be common multiplexed molecular assay (either Immucor PreciseType population variants or variants that have not previously HEA molecular BeadChip test, herein referred to as “HEA” been associated with established blood group alleles or or the RHD Molecular BeadChip test). phenotypes. The HEA test predicts antigen phenotype for RH (C, Here we investigate the potential application for c, E, e, V, VS), KEL (K, k, Kpa,Kpb,Jsa,Jsb), FY (Fya,Fyb, blood group genotyping using targeted exome sequencing GATA, Fyx), JK (Jka,Jkb), MNS (M, N, S, s, Uvar, Uneg), LU to resolve complex cases, where sequencing was (Lua,Lub), DO (Doa,Dob,Hy,Joa), LW (LWa,LWb), DI (Dia, b a b 14 requested by the Australian Red Cross Blood Service Red Di ), CO (Co ,Co ), and SC (Sc1, Sc2). The RHD test uti- Cell Reference Laboratory (RCRL). These requests were lizes 35 genetic markers associated with D phenotypes to 15 made either where serology typing was discordant to SNP make RHD variant calls. genotyping or where confirmation was needed to support antigen typing. MPS library preparation and sequencing We prepared DNA libraries for sequencing using a MATERIALS AND METHODS sequencing panel (TruSight One [TSO], Illumina) as per manufacturer’s instructions. The TSO panel amplifies RCRL serology investigations exons and untranslated regions for 4813 clinically signifi- Samples referred to investigate the D status were first cant genes, including the 41 genes related to 33 blood tested with two anti-D antisera: one, Epiclone-2 anti-D group systems and genes for two transcription factors, (IgM) (Seqirus) that does not detect most partial D and KLF1 and GATA1 (Table S1, available as supporting infor- weak D types, and another, anti-D (monoclonal blend) mation in the online version of this paper). The genes for Gamma-clone (Immucor) that does detect most partial D three (JMH, VEL, and XG) of the 36 blood group systems and weak D types. Samples were further tested with were excluded by the manufacturer of the kit. MPS was ALBAclone advanced partial Rh(D) typing kit (Alba Biosci- performed on the MiSeq sequencing platform (Illumina) ence) as per the manufacturer’s instruction. to generate paired-end 150-bp reads as described 13 Phenotyping was performed, where possible, with previously. commercially available antisera. When commercial anti- sera were not available, in-house antisera was used with Data analysis the optimum technique for that particular antiserum. FASTQ files were generated by the MiSeq system on- Where weak antigen expression was suspected, adsorption- instrument software and exported to computer software Volume 58, February 2018 TRANSFUSION 285 SCHOEMAN ET AL. (CLC Genomics Workbench software, Version 8.5, QIA- GEN) for secondary
Load more

Recommended publications
  • Genetic Screening for the Vel- Phenotype Circumvents Difficult Serological Screening Due to Variable Vel Expression Levels
    UvA-DARE (Digital Academic Repository) Genetic basis of rare blood group variants Wigman, L. Publication date 2013 Link to publication Citation for published version (APA): Wigman, L. (2013). Genetic basis of rare blood group variants. General rights It is not permitted to download or to forward/distribute the text or part of it without the consent of the author(s) and/or copyright holder(s), other than for strictly personal, individual use, unless the work is under an open content license (like Creative Commons). Disclaimer/Complaints regulations If you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: https://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. UvA-DARE is a service provided by the library of the University of Amsterdam (https://dare.uva.nl) Download date:28 Sep 2021 Chapter 6 Genetic screening for the Vel- phenotype circumvents difficult serological screening due to variable Vel expression levels Lonneke Haer-Wigman1 Shabnam Solati1 Aïcha Ait Soussan1 Erik Beckers2 Pim van der Harst3 Marga van Hulst-Sundermeijer4 Peter Ligthart1 Dick van Rhenen5 Hein Schepers3 Tamara Stegmann1 Masja de Haas1 C. Ellen van der Schoot1 1 Sanquin Research, Amsterdam and Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, Amsterdam, the Netherlands 2 Maastricht University Medical Centre+, Maastricht, The Netherlands 3 University Medical Center Groningen, University of Groningen, Groningen, The Netherlands 4 Sanquin Diagnostic Services, Amsterdam, The Netherlands 5 Sanquin Blood Bank, Rotterdam, Netherlands Manuscript under preparation Chapter 6 Abstract Background: Serological determination of the Vel- phenotype is challenging due to variable Vel expression levels.
    [Show full text]
  • Fcγr Binding and ADCC Activity of Human Igg Allotypes
    fimmu-11-00740 May 4, 2020 Time: 17:25 # 1 ORIGINAL RESEARCH published: 06 May 2020 doi: 10.3389/fimmu.2020.00740 FcgR Binding and ADCC Activity of Human IgG Allotypes Steven W. de Taeye1,2*†, Arthur E. H. Bentlage2, Mirjam M. Mebius3, Joyce I. Meesters3, Suzanne Lissenberg-Thunnissen2, David Falck4, Thomas Sénard4, Nima Salehi1, Manfred Wuhrer4, Janine Schuurman3, Aran F. Labrijn3, Theo Rispens1‡ and Gestur Vidarsson2‡ 1 Sanquin Research and Landsteiner Laboratory, Department of Immunopathology, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands, 2 Sanquin Research and Landsteiner Laboratory, Department of Experimental Edited by: Immunohematology, Amsterdam UMC, University of Amsterdam, Amsterdam, Netherlands, 3 Genmab, Utrecht, Eric O. Long, Netherlands, 4 Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, Netherlands National Institute of Allergy and Infectious Diseases (NIAID), United States Antibody dependent cellular cytotoxicity (ADCC) is an Fc-dependent effector function Reviewed by: of IgG important for anti-viral immunity and anti-tumor therapies. NK-cell mediated Amy W. Chung, ADCC is mainly triggered by IgG-subclasses IgG1 and IgG3 through the IgG-Fc- The University of Melbourne, Australia receptor (FcgR) IIIa. Polymorphisms in the immunoglobulin gamma heavy chain gene Geoffrey Thomas Hart, University of Minnesota Twin Cities, likely form a layer of variation in the strength of the ADCC-response, but this has United States never been studied in detail. We produced all 27 known IgG allotypes and assessed *Correspondence: FcgRIIIa binding and ADCC activity. While all IgG1, IgG2, and IgG4 allotypes behaved Steven W. de Taeye [email protected]; similarly within subclass, large allotype-specific variation was found for IgG3.
    [Show full text]
  • Blood Bank I D
    The Osler Institute Blood Bank I D. Joe Chaffin, MD Bonfils Blood Center, Denver, CO The Fun Just Never Ends… A. Blood Bank I • Blood Groups B. Blood Bank II • Blood Donation and Autologous Blood • Pretransfusion Testing C. Blood Bank III • Component Therapy D. Blood Bank IV • Transfusion Complications * Noninfectious (Transfusion Reactions) * Infectious (Transfusion-transmitted Diseases) E. Blood Bank V (not discussed today but available at www.bbguy.org) • Hematopoietic Progenitor Cell Transplantation F. Blood Bank Practical • Management of specific clinical situations • Calculations, Antibody ID and no-pressure sample questions Blood Bank I Blood Groups I. Basic Antigen-Antibody Testing A. Basic Red Cell-Antibody Interactions 1. Agglutination a. Clumping of red cells due to antibody coating b. Main reaction we look for in Blood Banking c. Two stages: 1) Coating of cells (“sensitization”) a) Affected by antibody specificity, electrostatic RBC charge, temperature, amounts of antigen and antibody b) Low Ionic Strength Saline (LISS) decreases repulsive charges between RBCs; tends to enhance cold antibodies and autoantibodies c) Polyethylene glycol (PEG) excludes H2O, tends to enhance warm antibodies and autoantibodies. 2) Formation of bridges a) Lattice structure formed by antibodies and RBCs b) IgG isn’t good at this; one antibody arm must attach to one cell and other arm to the other cell. c) IgM is better because of its pentameric structure. P}Chaffin (12/28/11) Blood Bank I page 1 Pathology Review Course 2. Hemolysis a. Direct lysis of a red cell due to antibody coating b. Uncommon, but equal to agglutination. 1) Requires complement fixation 2) IgM antibodies do this better than IgG.
    [Show full text]
  • Introduction to the Rh Blood Group.Pdf
    Introduction to the Rhesus Blood Group Justin R. Rhees, M.S., MLS(ASCP)CM, SBBCM University of Utah Department of Pathology Medical Laboratory Science Program Objectives 1. Describe the major Rhesus (Rh) blood group antigens in terms of biochemical structure and inheritance. 2. Describe the characteristics of Rh antibodies. 3. Translate the five major Rh antigens, genotypes, and haplotypes from Fisher-Race to Wiener nomenclature. 4. State the purpose of Fisher-Race, Wiener, Rosenfield, and ISBT nomenclatures. Background . How did this blood group get its name? . 1937 Mrs. Seno; Bellevue hospital . Unknown antibody, unrelated to ABO . Philip Levine tested her serum against 54 ABO-compatible blood samples: only 13 were compatible. Rhesus (Rh) blood group 1930s several cases of Hemolytic of the Fetus and Newborn (HDFN) published. Hemolytic transfusion reactions (HTR) were observed in ABO- compatible transfusions. In search of more blood groups, Landsteiner and Wiener immunized rabbits with the Rhesus macaque blood of the Rhesus monkeys. Rhesus (Rh) blood group 1940 Landsteiner and Wiener reported an antibody that reacted with about 85% of human red cell samples. It was supposed that anti-Rh was the specificity causing the “intragroup” incompatibilities observed. 1941 Levine found in over 90% of erythroblastosis fetalis cases, the mother was Rh-negative and the father was Rh-positive. Rhesus macaque Rhesus (Rh) blood group Human anti-Rh and animal anti- Rh are not the same. However, “Rh” was embedded into blood group antigen terminology. The
    [Show full text]
  • Blood Group Genomics
    BLOOD GROUP GENOMICS Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of 36 blood group alleles among 396 Thai samples reveals region-specific variants Philaiphon Jongruamklang,1 Christoph Gassner,2 Stefan Meyer,2 Aksarakorn Kummasook,3 Marion Darlison,1 Chayanun Boonlum,4 Surin Chanta,5 Beat M. Frey,2 Martin L. Olsson,1,6* and Jill R. Storry 1,6* lood group antigen polymorphism shows great BACKGROUND: Blood group phenotype variation has variation in different world populations. The been attributed to potential resistance to pathogen reason for this is not completely understood; invasion. Variation was mapped in blood donors from however, it has been attributed to both Lampang (northern region) and Saraburi (central region), B Thailand, where malaria is endemic. The previously unknown blood group allele profiles were characterized ABBREVIATIONS: MALDI-TOF MS 5 matrix-assisted laser and the data were correlated with phenotypes. The high desorption/ionization time-of-flight mass spectrometry; PCR- incidence of the Vel-negative phenotype previously ASP 5 polymerase chain reaction with allele-specific reported in Thais was investigated. primers; SNP(s) 5 single nucleotide polymorphism(s). STUDY DESIGN AND METHODS: DNA from 396 From 1Hematology and Transfusion Medicine, Department of blood donors was analyzed by matrix-assisted laser Laboratory Medicine, Lund University, Lund, Sweden; desorption/ionization–time-of-flight mass spectrometry. 2Molecular Diagnostics & Research (MOC), Blood Transfusion Outliers were investigated by serology and DNA Service Zurich,€ Zurich-Schlieren,€ Switzerland; 3Department of sequencing. Allele discrimination assays for SMIM1 Medical Technology, School of Allied Health Sciences, rs1175550A/G and ACKR1 rs118062001C/T were University of Phayao, Phayao, Thailand; 4Transfusion Medicine, performed and correlated with antigen expression.
    [Show full text]
  • Comprehensive Red Blood Cell and Platelet Antigen Prediction from Whole Genome Sequencing: Proof of Principle
    BLOOD GROUP GENOMICS Comprehensive red blood cell and platelet antigen prediction from whole genome sequencing: proof of principle William J. Lane,1,2 Connie M. Westhoff,3 Jon Michael Uy,1 Maria Aguad,1 Robin Smeland-Wagman,1 Richard M. Kaufman,1 Heidi L. Rehm,1,2,4,5 Robert C. Green,2,5,6 and Leslie E. Silberstein7 for the MedSeq Project* rediction of red blood cell (RBC) and platelet BACKGROUND: There are 346 serologically defined (PLT) antigens using DNA assays has the poten- red blood cell (RBC) antigens and 33 serologically tial to augment or replace traditional serologic defined platelet (PLT) antigens, most of which have antigen typing in many situations. DNA-based known genetic changes in 45 RBC or six PLT genes that P typing methods are more easily automated, amenable to correlate with antigen expression. Polymorphic sites multiplexing, and do not require expensive and some- associated with antigen expression in the primary times difficult to obtain serologic immunoglobulin literature and reference databases are annotated according to nucleotide positions in cDNA. This makes antigen prediction from next-generation sequencing data ABBREVIATIONS: CDS 5 coding DNA sequence; NGS 5 challenging, since it uses genomic coordinates. next-generation sequencing; SNP(s) 5 single-nucleotide STUDY DESIGN AND METHODS: The conventional polymorphism(s); WGS 5 whole genome sequencing. cDNA reference sequences for all known RBC and PLT From the 1Department of Pathology, the 6Division of Genetics, genes that correlate with antigen expression were Department of Medicine, and the 7Division of Transfusion aligned to the human reference genome. The alignments Medicine, Department of Pathology, Brigham and Women’s allowed conversion of conventional cDNA nucleotide Hospital; and 2Harvard Medical School, Boston, Massachusetts; positions to the corresponding genomic coordinates.
    [Show full text]
  • Clinical Significance of Antibodies to Antigens in the Raph, John Milton
    R EVIEW Proceedings from the International Society of Blood Transfusion Working Party on Immunohaematology, Workshop on the Clinical Significance of Red Blood Cell Alloantibodies, September 2, 2016, Dubai Clinical significance of antibodies to antigens in the Raph, John Milton Hagen, I, Globoside, Gill, Rh-associated glycoprotein, FORS, JR, LAN, Vel, CD59, and Augustine blood group systems M. Moghaddam and A.A. Naghi This article reviews information on the clinical significance and 6 shared missense mutation c.511C>T (p.Argl71Cys) as of antibodies to antigens in the Raph, John Milton Hagen, I, well as a synonymous single-nucleotide mutation (c.579A>G) Globoside, Gill, Rh-associated glycoprotein, FORS, JR, LAN, Vel, and had no clinical features. Although the CD151 protein is CD59, and Augustine blood group systems. Antibodies to many of the antigens in these groups are rarely encountered because of the critical to cell adhesion and signaling and is implicated in high prevalence of the associated antigens in most populations. cancer progression, its significance in transfusion medicine is For many of these antibodies, the clinical significance—that is, limited to only one report of a hemolytic transfusion reaction the potential to cause reduced survival of transfused antigen- 3 positive red blood cells or a transfusion reaction (e.g., anti-P, (HTR). Least-incompatible RBC units should be selected anti-Jra, and anti-Lan), and/or hemolytic disease of the fetus and for transfusion to patients with anti-MER2.2 No information newborn (e.g., anti-RHAG4 and anti-Vel)—has been documented. on anti-MER2 causing hemolytic disease of the fetus and For other antibodies, their prevalence is so rare that information newborn (HDFN) is available.4 on the clinical significance of their antibodies is not available (e.g., anti-FORS1).
    [Show full text]
  • This Issue of Immunohematology Is Supported by a Contribution From
    Journal of Blood Group Serology and Molecular Genetics VOLUME 33, N UMBER 4, 2017 This issue of Immunohematology is supported by a contribution from Grifols Diagnostics Solutions, Inc. Dedicated to advancement and education in molecular and serologic immunohematology Immunohematology Journal of Blood Group Serology and Molecular Genetics Volume 33, Number 4, 2017 CONTENTS O RIGINAL R EP O RT 147 Assessment of common red blood cell pretreatments to yield an accurate serologic antigen phenotype compared with genotype- predicted phenotype T. Horn, J. Hamilton, J. Kosanke, V.W. Hare, W. Kluver, W. Beres, S. Nance, and M.A. Keller C ASE R EP O RT 152 Anti-Vel alloimmunization and severe hemolytic disease of the fetus and newborn K.J. Moise Jr., Y. Morales, M.F. Bertholf, S.N. Rossmann, and Y. Bai S ER O LO GI C M ETH O D R EVIEW 155 Separation of multiple antibodies by adsorption with allogeneic red blood cells E.M. Ekema O RIGINAL R EP O RT 159 Hemovigilance and the Notify Library B.I. Whitaker, D.M. Strong, M.J. Gandhi, and E. Petrisli O RIGINAL R EP O RT 165 Clinical and laboratory profile of anti-M D. Basu, S. Basu, M. Reddy, K. Gupta, and M. Chandy S ER O LO GI C M ETH O D R EVIEW 170 Dithiothreitol treatment of red blood cells C.B. Bub C omm UNI C AT I O NS 173 Thank You S. Nance and C. Flickinger 174 175 183 187 191 E R R AT U M A NN O UN C E M ENTS A DVERTISE M ENTS I NSTRU C TI O NS S UBS C RIPTI O N F O R A UTH O RS I NF O R M AT I O N E DITO R - IN -C HIEF E DITO RIAL B OARD Sandra Nance, MS, MT(ASCP)SBB Philadelphia, Pennsylvania Patricia Arndt, MT(ASCP)SBB Geralyn M.
    [Show full text]
  • Mission Impossible with Anti-Vel
    Mission Impossible with Anti-Vel BBTS September 2014 Stephanie Stone, Sheffield Teaching Hospitals Helen Kilgallon, NHSBT Sheffield 74 yr old female Presented to ENT May 2013: 4/52 bilateral nasal obstruction and intermittent epistaxis No B symptoms Past medical history: - Metallic mitral valve replacement 1974: Warfarin INR 3-4 - Dilated left atrium, AF, Pulmonary HTN - Angina, COPD - Type II DM, CKD stage 4, Gout, Bullous pemphygoid Social history - Retired, lives alone - 4 daughters – first in 1957 - Ex smoker - No regular ETOH - Exercise tolerance 25-40 yards - ECOG performance status 2 - Independent of activities of daily living Drug history - No known drug allergies – mild allergic reaction to blood component 2012 - Warfarin - Digoxin 125mcg od - Furosemide 40mg bd - Pravastatin 40mg on - Candesartan 8mg od - Linagliptin 5mg od - Quinine 300mg od - Betahistine 8mg tds Examination and investigations www.trialx.com Fibre optic examination: large post nasal mass – Fine needle aspiration taken 2cm cervical LN. No other palpable LN/hepatosplenomegaly/masses Hb 97 Mcv 92 Platelets 287 Wbc 6.9 Lymph 0.6 Reticulocytes 248 B12 394 Folate 8.0 Ferritin 252 Bil 10 Alt 40 Alb 44 LDH 708 DAT –ve Haptoglobin normal Immunoglobulins normal HIV/Hep B/C serology normal Bone marrow: normal Echo: good LV function, dilated LA, moderate TR, increased PA pressure CT neck, chest, abdomen and pelvis R-sided post nasal mass CT chest, abdomen and pelvis Multiple cervical and mediastinal lymph nodes Fine needle aspiration High grade cell Non Hodgkins Lymphoma:
    [Show full text]
  • Red Blood Cell Antigen Genotyping
    Red Blood Cell Antigen Genotyping Testing is useful in determining allelic variants predicting red blood cell (RBC) antigen phenotypes for patients with recent history of transfusion or with conflicting serological antibody results due to partial, variant, or weak expression antigens. Also Tests to Consider useful as an aid in management of hemolytic disease of the fetus and newborn (HDFN). Typical Testing Strategy Disease Overview Phenotype Testing Evaluates specific RBC antigen presence by serology Prevalence and/or Incidence Results can aid in selecting antigen negative RBC units Erythrocyte alloimmunization occurs in up to 58% of sickle cell patients, up to 35% in other transfusion-dependent patients, and in approximately 0.8% of all pregnant Antigen Testing, RBC Phenotype women. Extended 0013020 Method: Hemagglutination Serological testing includes K, Fya, Fyb, Jka, Symptoms Jkb, S, s (k, cellano, testing performed if indicated) to assess maternal or paternal RBC Transfusion reactions or HDFN can occur due to alloimmunization: phenotype status. Antigen Testing, Rh Phenotype 0013019 Intravascular hemolysis: hemoglobinuria, jaundice, shock Extravascular hemolysis: fever and chills Method: Hemagglutination HDFN: fetal hemolytic anemia, hepatosplenomegaly, jaundice, erythroblastosis, Antigen testing for D, C, E, c, and e to assess neurological damage, hydrops fetalis maternal, paternal, or newborn Rh phenotype status Clinical presentation is variable and dependent upon the specific antibody and recipient factors Genotype Testing May help
    [Show full text]
  • Anti-Rhd Immunoglobulin in the Treatment of Immune Thrombocytopenia
    REVIEW Anti-RhD immunoglobulin in the treatment of immune thrombocytopenia Eric Cheung Abstract: Immune thrombocytopenia (ITP) is an acquired bleeding autoimmune disorder Howard A Liebman characterized by a markedly decreased blood platelet count. The disorder is variable, frequently having an acute onset of limited duration in children and a more chronic course in adults. Jane Anne Nohl Division of Hematology A number of therapeutic agents have demonstrated effi cacy in increasing the platelet counts in and Center for the Study of Blood both children and adults. Anti-RhD immunoglobulin (anti-D) is one such agent, and has been Diseases, University of Southern California-Keck School of Medicine, successfully used in the setting of both acute and chronic immune thrombocytopenia. In this Los Angeles, CA, USA report we review the use of anti-D in the management of ITP. While the FDA-approved dose of 50 mg/kg has documented effi cacy in increasing platelet counts in approximately 80% of children and 70% of adults, a higher dose of 75 μg/kg has been shown to result in a more rapid increase in platelet count without a greater reduction in hemoglobin. Anti-D is generally inef- fective in patients who have failed splenectomy. Anti-RhD therapy has been shown capable of delaying splenectomy in adult patients, but does not signifi cantly increase the total number of patients in whom the procedure can be avoided. Anti-D therapy appears to inhibit macrophage phagocytosis by a combination of both FcR blockade and infl ammatory cytokine inhibition of For personal use only. platelet phagocytosis within the spleen.
    [Show full text]
  • Prevalence of ABO, Rhd and Other Clinically Significant Blood Group Antigens Among Blood Donors at Tertiary Care Center, Gwalior
    ORIGINAL ARTICLE Bali Medical Journal (Bali Med J) 2020, Volume 9, Number 2: 437-443 P-ISSN.2089-1180, E-ISSN.2302-2914 Prevalence of ABO, RhD and other clinically ORIGINAL ARTICLE significant Blood Group Antigens among blood donors at tertiary care center, Gwalior Published by DiscoverSys CrossMark Doi: http://dx.doi.org/10.15562/bmj.v9i2.1779 Shelendra Sharma, Dharmesh Chandra Sharma,* Sunita Rai, Anita Arya, Reena Jain, Dilpreet Kaur, Bharat Jain Volume No.: 9 ABSTRACT Issue: 2 Background: Among all blood group systems and antigens, ABO and of them 1000 randomly selected donor samples were processed for D antigens of the Rh blood group system are of primary importance, complete Rh, Kell, Duffy and Kidd blood grouping. hence included in routine blood grouping and transfusion. Other blood Result: ABO group pattern was; A - 22.56%, B - 36.52%, AB - 9.8% group systems and antigens that are important in multiparous women, and O -31.12 %, while RhD status was RhD+ 90.99% and RhD- 9.01%. First page No.: 437 multi-transfused patients and hemolytic disease of newborn are Kell, Prevalence of Rh phenotypes was DCCee 43%, DCcee 33%, DCcEe 10%, Rh, Duffy, and Kidd. dccee 6.5%, DccEe 4.5%, Dccee 1%, DCCEe 0.3% and dccEe 0.2%. Aims and Objectives: To know the pattern of ABO, RhD and other Kell phenotype was K-k+ 95.5%, K+k+ 4.5%, while K+ k- and K- clinically significant major antigens of Rh, Kell, Duffy and Kidd blood k- phenotypes were not encountered in this study. Duffy phenotypes P-ISSN.2089-1180 group systems as well as evaluation of phenotypes, genotypes and were Fya+ Fyb+ 47.5%, Fya+ Fyb- 35%, Fya- Fyb+17% and Fya- Fyb- gene frequency of related antigens among blood donors in Gwalior 0.5%.
    [Show full text]