Comprehensive Platelet Disorder Panel

References Mumford AD, Nisar S et al. 2013. Platelet dysfunction associated with the novel Trp29Cys thromboxane A₂ receptor variant. J Thromb Haemost. Mar;11(3):547-54. Albers CA, Paul DS et al. 2012. Compound inheritance of a low-frequency regulatory SNP and a rare null mutation in exon-junction complex subunit RBM8A Noris P, Biino G et al. 2014. Platelet diameters in inherited thrombocytopenias: causes TAR syndrome. Nat Genet. 2012 Feb 26; 44(4):435-9. analysis of 376 patients with all known disorders. Blood. Aug 7; 124(6):e4-e10. Comprehensive Ali S, Ghosh K et al. 2016. Congenital macrothrombocytopenia is a heterogeneous Noris P, Favier R et al. 2013. ANKRD26-related thrombocytopenia and myeloid disorder in India. Haemophilia. Jul; 22(4):570-82. malignancies. Blood .122:1987-1989. Balduini CL, Pecci A, Savoia A. 2011. Recent advances in the understanding and Nurden AT, Nurden P. 2011. Advances in our understanding of the molecular basis management of MYH9-related inherited thrombocytopenias. Br J Haematol. of disorders of platelet function. J Thromb Haemost. 9 Suppl 1:76-91. Jul;154(2):161-174. Nurden AT, Nurden P. 2015. Inherited disorders of platelet function: selected Platelet Disorder Balduini CL, Melazzini F, Pecci A. 2017. Inherited thrombocytopenias-recent updates. J Thromb Haemost. Volume 13(Suppl1): S2- S9. advances in clinical and molecular aspects. Platelets. Jan;28(1):3-13. Ok Bozkaya I, Yarali N et al. Severe Clinical Course in a Patient with Congenital Bastida JM, Del Rey M et al. 2016. Wiskott-Aldrich syndrome in a child presenting Amegakaryocytic Thrombocytopenia Due to a Missense Mutation of the c-MPL with macrothrombocytopenia. Platelets.Nov 25:1-4. Gene. Turk J Haematol.Jun; 32(2): 172-4. Panel Bottega R, Marconi C et al. 2015. ACTN1-related thrombocytopenia: identification Pagel J, Beutel K et al. 2012. Distinct mutations in STXBP2 are associated of novel families for phenotypic characterization. Blood. Jan 29;125(5):748-50. with variable clinical presentations in patients with familial hemophagocytic lymphohistiocytosis type 5 (FHL5). Blood. 119:6016-6024. Cox K, Price V, Kahr WHA. 2011. Inherited platelet disorders: a clinical approach to diagnosis and management. Expert Rev Hematol. Aug;4(4):455-72. Paterson AD, Rommens JM, et al. 2010. Persons with Quebec platelet disorder have a tandem duplication of PLAU, the urokinase plasminogen activator gene. difficult to interpret, and typically require immediate testing on Faioni EM, Razzari C et al. 2014. Bleeding diathesis and gastro-duodenal ulcers Blood.115:1264-6. BloodCenter of Wisconsin offers a specifically in inherited cytosolic phospholipase-A2 alpha deficiency. Thromb Haemost. fresh patient platelets due to limited sample stability. Advances in Dec;112(6):1182-9. Rao AK. 2013. Inherited platelet function disorders: overview and disorders of designed Comprehensive Platelet Disorder granules, secretion, and signal transduction. Hematol Oncol Clin North Am. Jun; genetic testing through next-generation sequencing allows for Favier R, Raslova H. 2015. Progress in understanding the diagnosis and molecular 27(3):585-611. Panel (test code 4830) optimized for detection identification of underlying genetic defects and for distinguishing genetics of macrothrombocytopenias. Br J Haematol. Sep;170 (5):626-39. Rehm HL, Bale SJ et al. 2013. Working Group of the American College of Medical of germline variants in 43 genes known to inherited platelet disorder cases from immune thrombocytopenia. Freson K, Wijgaerts A, Van Geet C. 2014. Update on the causes of platelet disorders Genetics and Genomics Laboratory Quality Assurance Committee. ACMG clinical cause platelet function disorders and/or Accurate diagnosis provides information about the phenotype and functional consequences. John Wiley & Sons Ltd, Int. Jnl. Lab. Hem. 36, laboratory standards for next-generation sequencing. Genet Med.15:733-747. and prognosis, guides medical management decisions, assists 313–325. Richards S, Aziz N A et al.2015. Standards and guidelines for the interpretation of inherited thrombocytopenia. with the identification of affected family members, and allows for Gresele P. 2014. Diagnosis of inherited platelet function disorders: guidance for the sequence variants: a joint consensus recommendation of the American College accurate genetic recurrence risk assessment. SSC of the ISTH. J Thromb and Haemost. 13:314-322. of Medical Genetics and Genomics and the Association for Molecular Pathology. Gunay-Aygun M, Falik-Zaccai TC et al. 2011. NBEAL2 is mutated in Gray platelet Genet Med.17:405-424. Platelet function disorders and inherited thrombocytopenia are a Variants in several genes known to cause syndromic or non- Syndrome and is required for biogenesis of platelet alpha-granules. Nat Genet. Savoia A. 2016. Molecular basis of Inherited thrombocytopenias. Clin Genet heterogeneous group of disorders which may have overlapping syndromic platelet disorders may be inherited in an autosomal Aug; 43(8): 732–734. Feb;89(2):154-62. clinical phenotypes, generally differentiated by platelet counts. recessive, autosomal dominant or X-linked recessive manner. More Hinckley J, Di Paola J. 2014. Genetic basis of congenital platelet disorders. Savoia A. 2016. Molecular basis of Inherited thrombocytopenias: an update. Inherited thrombocytopenia disorders are typically characterized common and rare types of syndromic or non-syndromic platelet Hematology Am Soc Hematol Educ Program. Dec5;(1): 3337-42. Current Opinion in Hematology. Sep;23(5):486-492. by platelet counts less than 150,000/uL, but often can vary with disorders will be identified with this panel, including MYH9-related Israels SJ, El-Ekiaby M, Quiroga T, Mezzano D. 2010. Inherited disorders of platelet Kunishima Shinji S, Kobayashi R et al. 2009. Mutation of the β1-tubulin gene age, gender and ethnic background, while platelet function disorders, Glanzmann thrombasthenia, Bernard-Soulier syndrome, function and challenges to diagnosis of mucocutaneous bleeding. Haemophilia. associated with congenital macrothrombocytopenia affecting microtubule disorders are usually, but not always, associated with normal congenital amegakaryocytic thrombocytopenia, familial 16 (Suppl 5):152-9. assembly. Blood. 113:458-461. platelet counts and are caused by defects in platelet adhesion, platelet disorder with predisposition for acute myelogenous Johnson B, Lowe GC et al. 2016. Whole exome sequencing identifies genetic Songdej N, Rao AK. 2017. Inherited platelet dysfunction and hematopoietic glycoprotein expression, receptor function, signaling pathways, leukemia, ANKRD26-related thrombocytopenia, WAS-related variants in inherited thrombocytopenia with secondary qualitative function transcription factor mutations. Platelets Jan; 28(1):20-26. aggregation, cytoskeleton proteins, secretion, granular contents thrombocytopenia, Scott syndrome, gray platelet syndrome defects. Haematologica. 101(10):1170-1179. Stevenson WS, Rabbolini DJ et al. 2015. Paris-Trousseau thrombocytopenia is and abnormalities in procoagulant activity. Symptoms of platelet and others. Additional genes on this panel are associated with Kumar R, Kahr WHA. 2013. Congenital Thrombocytopenia: Clinical Manifestations, phenocopied by the autosomal recessive inheritance of a DNA-binding domain disorders may include purpura, petechiae, prolonged bleeding syndromes that have platelet disorders as a common finding Laboratory Abnormalities, and Molecular Defects of a Heterogeneous Group of mutation in FLI1. Blood. Oct 22;126(17); 2027-30. Conditions. Hematol Oncol Clin North Am. June; 27(3):465-94. from cuts, epistaxis, gum bleeding, excessive bleeding after among other non-hematologic features. Watson SP, Lowe M et al. 2013. Genotyping and phenotyping of platelet function surgery, hemoptysis, hematuria and menorrhagia in women. Kunicki TJ, Williams SA, Nugent DJ. 2012. Genetic variants that affect platelet disorders. J Thromb Haemost. 11(suppl.1):351-363. This panel evaluates for single nucleotide variants and small function. Current Opin Hematol.19:371-9. Severe platelet disorders can present in the newborn period, deletions and duplications, which are most commonly responsible Westbury SK, Mumford AD. 2016. Genomics of platelet disorders. Hemophilia. while mild thrombocytopenia may remain undiagnosed until Lentaigne C, Freson K et al. 2016. Inherited platelet disorders: toward DNA-based 22(Suppl.5): 20-24. for genetic disease. However, large deletions and duplications, diagnosis. Blood. 127(23):2814-2823. incidental detection of thrombocytopenia on routine blood also referred to as copy number variants (CNV), are a known cause Zhang My, Churpek JE et al. 2015. Germline ETV6 mutations in familial testing in adulthood. Some inherited platelet disorders have Manchev VT, Hilpert M et al. 2014. A new form of macrothrombocytopenia thrombocytopenia and hematologic malignancy. Nat Genet. Feb; 47(2):180-185. of genetic disorders, but can escape detection by next-generation induced by a germ-line mutation in the PRKACG gene. Blood. Oct 16; only hematologic manifestations, such as differences in platelet sequence analysis. Further testing with BloodCenter of Wisconsin 124(16):2554-63. Zhou Y, Zhang J. 2014. Arthrogryposis–renal dysfunction–cholestasis (ARC) size or distinctive granulocyte inclusions, while other syndromic syndrome: from molecular genetics to clinical features. Italian J Pediatrics. 40:77. custom designed, high density gene-focused array, aCGH Maclachlan A, Watson SP, Morgan NV. 2017. Inherited platelet disorders: Insight types present with additional non-hematologic manifestations. Deletion/Duplication Analysis, allows for the possible detection from platelet genomics using next-generation sequencing. Platelets. Jan ;28(1);14- Certain types of platelet disorders cause predisposition to acute 19. of large deletions and duplications within a single exon of a given myelogenous leukemia or myelodysplastic syndromes. gene, encompassing one or more exons, or affecting an entire Misdiagnosis of platelet disorders can result in inappropriate gene. This testing may be warranted when results of sequence therapies and inadequate surveillance for additional medical analysis do not fully explain a clinical phenotype, or when a complications, underscoring the importance of accurate suspected disorder is known to be caused by diagnosis. The diagnosis may be difficult to establish based solely on functional studies, especially in patients with milder disorders, as these assays are often technically challenging,

© Copyright 2017 r0818 BloodCenter of Wisconsin, Inc. , Part of Versiti. All rights reserved. A non-profit 503(c). deletions or duplications. Specifically, the Quebec Platelet gene in the Comprehensive Platelet Disorder Panel, including Shipping requirements CPT Codes/Billing/Turnaround time Disorder is associated with a heterozygous 78-kb tandem the clinical phenotype, OMIM numbers and inheritance pattern. Ship on an ice pack or at room Test Code: 4830 duplication of the PLAU gene, which will be detected by aCGH temperature. Protect from freezing. and not by the Comprehensive Platelet Disorder Panel. Please CPT codes: 81404, 81406, 81479 Place the specimen and the requisition refer to the aCGH Deletion/Duplication Analysis test description into plastic bags and seal. Insert into Turnaround time: 21 days for more information about specific genes included in this array. a Styrofoam container, seal and place The CPT codes provided are subject to change as more Refer to the table insert for further information about each into a sturdy cardboard box, and information becomes available. CPT codes are provided only as tape securely. Ship the package in guidance to assist clients with billing. compliance with your overnight carrier D P For additional information related to shipping, billing or pricing, guidelines. Label with the following please contact, BloodCenter Client Services: (414) 937-6396 or address: Indications for testing Assay sensitivity and limitations 800-245-3117, Option 1, or [email protected]. Client Services/Diagnostic Laboratory Comprehensive Platelet Disorder Panel: The analytical sensitivity of this test is >99% for single nucleotide BloodCenter of Wisconsin • Clarification and/or confirmation of diagnosis in a patient with changes and insertions and deletions of less than 20 bp. This 638 N. 18th St. clinical findings of a platelet disorder or an associated genetic assay does not detect large deletions or duplications (>20 bp) Milwaukee, WI 53233 syndrome when patient’s history suggests multiple possible or deletions, duplications or variants that are outside the regions platelet disorders sequenced. To order analysis of copy number variants at the exon or gene level, please refer to the aCGH Deletion/Duplication Required forms • Identification of carriers with family history of an unspecified Analysis test, if available, or contact Client Services before placing Please complete all pages of the platelet disorder to provide accurate reproductive risk your order. assessment and genetic counseling requisition form. Clinical history (including patient’s ethnicity, clinical Single gene sequencing or custom gene panel: Reporting of results diagnosis, family history and relevant • Analysis of genes included in the Comprehensive Platelet laboratory findings) is necessary for Disorder Panel may also be ordered as a stand-alone single While this assay is designed to detect germline genetic optimal interpretation of genetic test gene sequencing test or custom panel (2-10 genes) as dictated variants associated with platelet function disorders, variants D results and recommendations. ClinicalP by the patient’s clinical and laboratory phenotype unrelated to the indication for testing, but with other clinical and laboratory history can either be and/or reproductive implications, may also be detected. A Targeted familial variant analysis: recorded on the requisition form or comprehensive database of gene-phenotype relationships listed clinical and laboratory reports can be • Targeted variant analysis for clinical diagnosis, carrier by gene name can be found at http://www.omim.org. submitted with the sample. identification or prenatal diagnosis can also be performed on Results are classified and reported in accordance with ACMG any gene in the panel when the pathogenic variant(s) is known next-generation sequencing standards. Variants predicted to in the family (test code: 4970) be pathogenic, likely pathogenic, and of uncertain significance For clinical questions about laboratory tests and test utilization will be reported; variants classified as likely benign or benign are support, contact BloodCenter Client Services: (414) 937-6396 or typically not reported but such data are available upon request. 800-245-3117, Option 1, to be directed to our genetic counselors Sequence variants are described using standard Human Genome and clinical support team. Variation Society (HGVS) nomenclature (http://hgvs.org).

Comprehensive Platelet Disorder Panel, Insert Comprehensive Platelet Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. Comprehensive Platelet Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. Gene Clinical Phenotype Phenotype/Gene Inheritance Gene Clinical Phenotype Phenotype/Gene Inheritance OMIM number OMIM number GP1BA Bernard-Soulier syndrome (BSS): mild to severe bleeding disorder due to absence 231200/606672 Autosomal Recessive MYH9 MYH9-related disorders: large platelets and thrombocytopenia at birth with variable 160775 or dysfunction of the platelet glycoprotein receptor Ib/V/IX complex with mild to later onset of non-hematologic manifestations including progressive sensorineural moderate thrombocytopenia, unusually large platelets and abnormal platelet function hearing loss, glomerulonephritis, presenile cataracts and elevation of liver enzymes. with absent or markedly reduced aggregation response to ristocetin. caused by MYH9-related disorders includes previously characterized disorders: homozygous or compound heterozygous pathogenic variants in GP1BA (606672), GP1BB May-Hegglin anomaly: thrombocytopenia, giant platelets, and Dohle body-like 155100/160775 (138720), or GP9 (173515). inclusions Bernard-Soulier syndrome (BSSA2): autosomal dominant (mono-allelic) form of BSS with 153670/606672 Autosomal Dominant Epstein syndrome: thrombocytopenia, giant platelets, nephritis, and deafness 153650/160775 mild thrombocytopenia, variable large platelets, and mild or no bleeding tendency, due Autosomal Dominant to specific heterozygous variants in GP1BA gene or rarely in GP1BB or GP9. Fechtner syndrome: thrombocytopenia, giant platelets, and Dohle body-like inclusions 153640/160775 in peripheral blood leukocytes, with nephritis, hearing loss, and eye abnormalities, Platelet-type von Willebrand disease (also known as pseudo-von Willebrand disease): 177820/606672 Autosomal Dominant mostly cataracts thrombocytopenia and mucosal bleeding due to dominant pathogenic variants in GP1BA that cause excessive binding of the GPIb-IX-V complex to von Willebrand factor. Sebastian syndrome: thrombocytopenia, giant platelets, and leukocyte inclusions 605249/160775 GP1BB 231200/138720 Autosomal Recessive composed of highly dispersed filaments and few ribosomes 153670/138720 Autosomal Dominant DNFA17: nonsyndromic progressive hearing loss with onset in childhood or later; 603622/160775 GP9 231200/173515 Autosomal Recessive hearing loss progressed from high frequency to moderate-severe deafness over time. 153670/173515 Autosomal Dominant NBEAL2 Gray platelet syndrome (GPS): large platelets that lack α-granules leading to mild 139090/614169 Autosomal Recessive GP6 Bleeding disorder, platelet-type 11 (BDPLT11): mild to moderate bleeding disorder 614201/605546 Autosomal Recessive to moderate bleeding tendency and moderate thrombocytopenia with increased due to glycoprotein VI deficiency with defective platelet activation and aggregation in development of myelofibrosis and splenomegaly. response to collagen. P2RY12 Bleeding disorder, platelet-type 8 (BDPLT8): mild to moderate mucocutaneous bleeding 609821/600515 Autosomal Recessive HOXA11 Radioulnar synostosis with amegakaryocytic thrombocytopenia 1 (RUSAT1): possible 605432/142958 Autosomal Dominant and excessive bleeding after surgery or trauma due to ADP receptor defect. aplastic anemia and proximal fusion of the radius and ulna. Thrombocytopenia symptoms may be present at birth and require bone marrow or umbilical cord stem-cell PLA2G4A Recurrent episodes of multiple complicated ulcers of the small intestine, platelet See reference/ Autosomal Recessive transplantation. dysfunction and globally decreased eicosanoid synthesis due to phospholipase A2 600522 deficiency (Faioni EM et al. 2014). HPS1 Hermansky-Pudlak syndrome (HPS): multisystem disorder characterized by 203300/604982 Autosomal Recessive PRKACG Bleeding disorder platelet-type 19 (BDPLT19): severe macrothrombocytopenia with 616176/176893 Autosomal Recessive HPS3 oculocutaneous albinism and platelet defects with reduced or absent delta granules on 614072/606118 Autosomal Recessive electron microscopy of platelets. Subtypes are caused by recessive pathogenic variants moderate-severe bleeding tendency due to defects in megakaryocyte proplatelet HPS4 in nine different genes and variable additional features including neutropenia and/or 614073/606682 Autosomal Recessive formation, platelet activation and cytoskeleton reorganization. HPS5 immune defects (HPS2), pulmonary fibrosis HPS1,( HPS2 and HPS4), or granulomatous 614074/607521 Autosomal Recessive RASGRP2 Bleeding disorder, platelet-type 18 (BDPLT18): mild bleeding with increased bleeding 615888/605577 Autosomal Recessive colitis (HPS1 and HPS4). HPS3 is associated with milder bleeding symptoms and minimal HPS6 614075/607522 Autosomal Recessive times and platelets showed reduced aggregation in response to ADP or epinephrine. hypopigmentation. RBM8A Thrombocytopenia-Absent Radius syndrome (TAR): characterized by bilateral absence 274000/605313 Autosomal Recessive ITGA2B Glanzmann thrombasthenia: mild to severe bleeding disorder with platelet aggregation 273800/607759 Autosomal Recessive of the radius with presence of thumbs, thrombocytopenia which may be congenital or abnormalities due to quantitative or qualitative defects of platelet glycoproteins IIb develop early in life and decreases with age. Other anomalies of the skeleton (upper and and /or IIIa. caused by homozygous or compound heterozygous recessive pathogenic lower limbs, ribs, and vertebrae), heart, and genitourinary system (renal anomalies and variants in ITGA2B or ITGB3. agenesis of uterus, cervix, and upper part of the vagina) can occur. Bleeding disorder, platelet-type 16 (BDPLT16): congenital macrothrombocytopenia 187800/607759 Autosomal Dominant RUNX1 Familial platelet disorder with associated myeloid malignancy (FPDMM): mild to 601399/151385 Autosomal Dominant associated with platelet anisocytosis with mild to or absent symptoms due to specific moderate thrombocytopenia, qualitative platelet defects and a predisposition to heterozygous dominant activating mutations in ITGA2B or ITGB3. ITGB3 273800/173470 Autosomal Recessive development of myeloid malignancies. STIM1 Stormorken syndrome: multisystem disorder characterized by mild bleeding tendency 185070/605921 Autosomal Dominant 187800/173470 Autosomal Dominant due to platelet dysfunction, thrombocytopenia, anemia, asplenia, tubular aggregate LYST Chediak-Higashi syndrome: multisystem disorder characterized by partial 214500/ 606897 Autosomal Recessive myopathy, congenital miosis, and ichthyosis, with variable headache or recurrent stroke- oculocutaneous albinism, immunodeficiency with neutropenia and giant neutrophil like episodes. granules, bleeding tendency due to absent, reduced or abnormal platelet-dense bodies, STXBP2 Familial hemophagocytic lymphohistiocytosis (FHL5): immune dysregulation with 613101/601717 Autosomal Recessive malignant lymphoma and varying neurologic problems. hypercytokinemia, defective function of natural killer cells, and macrophages that MPL Congenital amegakaryocytic thrombocytopenia (CAMT): type 1-onset in infancy with 604498/159530 Autosomal Recessive infiltrate multiple organs; characterized by variable age of onset and severity with fever, severe thrombocytopenia and progressing to pancytopenia; type II with transient chronic diarrhea, hepatosplenomegaly, pancytopenia, coagulation abnormalities and increases in platelet counts with onset of bone marrow failure at age 3 or later. CNS problems.

Comprehensive Platelet Disorder Panel, Insert Comprehensive Platelet Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. Comprehensive Platelet Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. Gene Clinical Phenotype Phenotype/Gene Inheritance Gene Clinical Phenotype Phenotype/Gene Inheritance OMIM number OMIM number TBXA2R Susceptibility to bleeding disorder platelet-type 13 (BDPLT13): susceptibility to mild 614009/188070 Autosomal Dominant ACTN1 Bleeding disorder, platelet-type 15 (BDPT15): macrothrombocytopenia with mild or 615193/102575 Autosomal Dominant mucocutaneous bleeding due to defective platelet thromboxane A2 receptor, with absent bleeding tendency. bleeding phenotype occurring only in the presence of a second variant in this gene or ANKRD26 Thrombocytopenia-2 (THC2): mild bleeding tendency with normal platelet function and 188000/610855 Autosomal Dominant another gene affecting platelet function. morphology with increased predisposition to hematologic myeloid malignancies. TUBB1 Congenital macrothrombocytopenia. 613112/612901 Autosomal Dominant ANO6 Scott syndrome: moderate bleeding disorder due to abnormality of platelet 262890/608663 Autosomal Recessive VIPAS39 Arthrogryposis, renal dysfunction and cholestasis syndrome (ARCS1 and 2): multisystem 613404/ 613401 Autosomal Recessive procoagulation activity with normal platelet count. Impaired exposure of (VIPAR) disorder characterized by early onset and limited survival with arthrogryposis, renal phosphatidylserine on the outer leaflet of platelet membrane. dysfunction and cholestasis with additional symptoms of ichthyosis, abnormal platelet Hermansky-Pudlak syndrome (HPS) : multisystem disorder characterized by 608233/603401 Autosomal Recessive VPS33B count and function, secondary infection, and cardiovascular anomalies 208085/608552 Autosomal Recessive AP3B1 (HPS2) oculocutaneous albinism and platelet defects with reduced or absent delta granules on WAS WAS-related disorders comprise a spectrum of disorders, not distinct entities, as BLOC1S3 electron microscopy of platelets. Subtypes are caused by recessive pathogenic variants 614077/609762 Autosomal Recessive clinical manifestations can vary, even within the same family: (HPS8) in nine different genes and variable additional features including neutropenia and/or Wiskott-Aldrich syndrome (WAS): profound thrombocytopenia, small platelet size, 301000/ 300392 immune defects (HPS2), pulmonary fibrosis (HPS1, HPS2 and HPS4), or granulomatous eczema and recurrent infections; increased risk for autoimmune disorders and BLOC1S6 colitis (HPS1 and HPS4). HPS3 is associated with milder bleeding symptoms and minimal 614171/604310 Autosomal Recessive lymphoma, absent or decreased intracellular WAS protein (WASP) detection in (HPS9) hypopigmentation. hematopoietic cells by flow cytometry or western blotting. X-linked Recessive DTNBP1 614076/607145 Autosomal Recessive X-linked neutropenia: congenital neutropenia; variable severity, infectious history, 300299/ 300392 (HPS7) myelodysplasia and increased risk for MDS and AML. Normal WAS protein (WASP) CYCS Thrombocytopenia-4 (THC4): bleeding tendency may be mild or absent with normal 612004/123970 Autosomal Dominant expression by flow cytometry or western blotting. platelet size and morphology. X-linked thrombocytopenia: thrombocytopenia, possibly intermittent, with small 313900 / 300392 ETV6 Thrombocytopenia-5 (THC5): onset is typically in early childhood with an increased 616216/600618 Autosomal Dominant platelet volume; variable severity of bleeding, autoimmune disease and malignancies, susceptibility to develop various hematologic and solid malignancies throughout life. variable WASP expression by flow cytometry or western blotting. FERMT3 Leukocyte integrity adhesion deficiency type III: increased bleeding symptoms, poor 612840/607901 Autosomal Recessive WIPF1 Wiskott-Aldrich syndrome-2 (WAS2): thrombocytopenia, recurrent infections, eczema, 614493/602357 Autosomal Recessive wound healing, normal/reduced platelets, bacterial infections and neutrophilia. B-cell and T-cell deficiency, impaired NK cell function; normal WASP sequence and mRNA; defective WASP expression. FLI1 Paris-Trousseau thrombocytopenia (TCPT): mild bleeding tendency with variable 188025/193067 Deletion syndrome thrombocytopenia, dysmorphic facies, abnormal giant alpha-granules in platelets and dysmegakaryopoiesis as part of contiguous gene deletion syndrome of 11q (Jacobsen syndrome). The PLAU gene is not included in this panel. For analysis of the Quebec Platelet Disorder associated with a heterozygous 78-kb tandem duplication of the PLAU gene, please order aCGH. Bleeding disorder, platelet-type, 21 (BDPLT21): moderate macrothrombocytopenia, 617443/ 193067 Autosomal Recessive/ abnormal giant alpha-granules in platelets, functional platelet disorder. Phenotype Autosomal Dominant caused by heterozygous and homozygous pathogenic variants. GATA1 GATA1-related cytopenia: thrombocytopenia and/or anemia ranging from mild to severe; may be associated with platelet dysfunction, mild β-thalassemia, neutropenia, and congenital erythropoietic porphyria (CEP). Thrombocytopenia present in infancy and anemia may range from mild to severe hydrops fetalis. Carrier females may have mild to moderate symptoms. X-linked dyserythropoietic anemia and thrombocytopenia (XLTDA): variable severity of 300367/305371 thrombocytopenia and dyserythropoietic anemia may be present. X-linked Recessive Thrombocytopenia with beta-thalassemia, X-linked (XLTT): variable thrombocytopenia, 314050/305371 splenomegaly, and unbalanced hemoglobin chain synthesis resembling that of beta- thalassemia minor. GFI1B Bleeding disorder, platelet-type 17 (Gray Platelet-like syndrome): variable 187900/604383 Autosomal Dominant bleeding symptoms due to disorder of platelet alpha granules with moderate macrothrombocytopenia and red cell anisopoikilocytosis.

Comprehensive Platelet Disorder Panel, Insert deletions or duplications. Specifically, the Quebec Platelet gene in the Comprehensive Platelet Disorder Panel, including Shipping requirements CPT Codes/Billing/Turnaround time Disorder is associated with a heterozygous 78-kb tandem the clinical phenotype, OMIM numbers and inheritance pattern. Ship on an ice pack or at room Test Code: 4830 duplication of the PLAU gene, which will be detected by aCGH temperature. Protect from freezing. and not by the Comprehensive Platelet Disorder Panel. Please CPT codes: 81404, 81406, 81479 Place the specimen and the requisition refer to the aCGH Deletion/Duplication Analysis test description into plastic bags and seal. Insert into Turnaround time: 21 days for more information about specific genes included in this array. a Styrofoam container, seal and place The CPT codes provided are subject to change as more Refer to the table insert for further information about each into a sturdy cardboard box, and information becomes available. CPT codes are provided only as tape securely. Ship the package in guidance to assist clients with billing. compliance with your overnight carrier D P For additional information related to shipping, billing or pricing, guidelines. Label with the following please contact, BloodCenter Client Services: (414) 937-6396 or address: Indications for testing Assay sensitivity and limitations 800-245-3117, Option 1, or [email protected]. Client Services/Diagnostic Laboratory Comprehensive Platelet Disorder Panel: The analytical sensitivity of this test is >99% for single nucleotide BloodCenter of Wisconsin • Clarification and/or confirmation of diagnosis in a patient with changes and insertions and deletions of less than 20 bp. This 638 N. 18th St. clinical findings of a platelet disorder or an associated genetic assay does not detect large deletions or duplications (>20 bp) Milwaukee, WI 53233 syndrome when patient’s history suggests multiple possible or deletions, duplications or variants that are outside the regions platelet disorders sequenced. To order analysis of copy number variants at the exon or gene level, please refer to the aCGH Deletion/Duplication Required forms • Identification of carriers with family history of an unspecified Analysis test, if available, or contact Client Services before placing Please complete all pages of the platelet disorder to provide accurate reproductive risk your order. assessment and genetic counseling requisition form. Clinical history (including patient’s ethnicity, clinical Single gene sequencing or custom gene panel: Reporting of results diagnosis, family history and relevant • Analysis of genes included in the Comprehensive Platelet laboratory findings) is necessary for Disorder Panel may also be ordered as a stand-alone single While this assay is designed to detect germline genetic optimal interpretation of genetic test gene sequencing test or custom panel (2-10 genes) as dictated variants associated with platelet function disorders, variants D results and recommendations. ClinicalP by the patient’s clinical and laboratory phenotype unrelated to the indication for testing, but with other clinical and laboratory history can either be and/or reproductive implications, may also be detected. A Targeted familial variant analysis: recorded on the requisition form or comprehensive database of gene-phenotype relationships listed clinical and laboratory reports can be • Targeted variant analysis for clinical diagnosis, carrier by gene name can be found at http://www.omim.org. submitted with the sample. identification or prenatal diagnosis can also be performed on Results are classified and reported in accordance with ACMG any gene in the panel when the pathogenic variant(s) is known next-generation sequencing standards. Variants predicted to in the family (test code: 4970) be pathogenic, likely pathogenic, and of uncertain significance For clinical questions about laboratory tests and test utilization will be reported; variants classified as likely benign or benign are support, contact BloodCenter Client Services: (414) 937-6396 or typically not reported but such data are available upon request. 800-245-3117, Option 1, to be directed to our genetic counselors Sequence variants are described using standard Human Genome and clinical support team. Variation Society (HGVS) nomenclature (http://hgvs.org).

Test method Specimen requirements This next generation sequencing assay analyzes 43 genes, Parental/Patient/Pediatric: 3-5 mL Whole Blood (EDTA tube, spanning the full coding regions plus a minimum 30bp of lavender top), 2-5 mL Bone Marrow (EDTA tube, lavender top), 3-4 non-coding DNA including intron-exon junctions, and to Buccal Swabs, or ≥1ug of DNA at ≥50ng/uL of High Quality DNA. approximately 200bp upstream of ANKRD26 coding region (5’ UTR). These targeted regions are captured by hybridization, Fetal: 7-15 mL Amniotic fluid, 5-10 mg Chorionic villi; back up amplified and sequenced by massively parallel sequencing. culture of amniocytes or chorionic villi is highly recommended. Regions will have a minimum coverage of 50x and those regions Cultured: Two T25 flasks cultured amniocytes or chorionic villi with less than 50 sequencing reads or low quality coverage are (2x106 minimum). Maternal Blood sample of 3-5 mL Whole Blood supplemented with Sanger sequencing. All regions are covered (EDTA tube, lavender top) is requested for all prenatal samples for by bi-directional analysis. Variants are identified by a customized maternal cell contamination studies. bioinformatics pipeline, analyzed and comprehensively If questions please contact the laboratory to discuss sample interpreted by our team of directors, scientists, and genetic requirements. counselors. All reported variants, including pathogenic, likely pathogenic, and variants of uncertain significance, are confirmed by Sanger sequencing. For prenatal testing, analysis of variable number tandem repeats (VNTR) is used to confirm results are not affected by maternal cell contamination. References Mumford AD, Nisar S et al. 2013. Platelet dysfunction associated with the novel Trp29Cys thromboxane A₂ receptor variant. J Thromb Haemost. Mar;11(3):547-54. Albers CA, Paul DS et al. 2012. Compound inheritance of a low-frequency regulatory SNP and a rare null mutation in exon-junction complex subunit RBM8A Noris P, Biino G et al. 2014. Platelet diameters in inherited thrombocytopenias: causes TAR syndrome. Nat Genet. 2012 Feb 26; 44(4):435-9. analysis of 376 patients with all known disorders. Blood. Aug 7; 124(6):e4-e10. Comprehensive Ali S, Ghosh K et al. 2016. Congenital macrothrombocytopenia is a heterogeneous Noris P, Favier R et al. 2013. ANKRD26-related thrombocytopenia and myeloid disorder in India. Haemophilia. Jul; 22(4):570-82. malignancies. Blood .122:1987-1989. Balduini CL, Pecci A, Savoia A. 2011. Recent advances in the understanding and Nurden AT, Nurden P. 2011. Advances in our understanding of the molecular basis management of MYH9-related inherited thrombocytopenias. Br J Haematol. of disorders of platelet function. J Thromb Haemost. 9 Suppl 1:76-91. Jul;154(2):161-174. Nurden AT, Nurden P. 2015. Inherited disorders of platelet function: selected Platelet Disorder Balduini CL, Melazzini F, Pecci A. 2017. Inherited thrombocytopenias-recent updates. J Thromb Haemost. Volume 13(Suppl1): S2- S9. advances in clinical and molecular aspects. Platelets. Jan;28(1):3-13. Ok Bozkaya I, Yarali N et al. Severe Clinical Course in a Patient with Congenital Bastida JM, Del Rey M et al. 2016. Wiskott-Aldrich syndrome in a child presenting Amegakaryocytic Thrombocytopenia Due to a Missense Mutation of the c-MPL with macrothrombocytopenia. Platelets.Nov 25:1-4. Gene. Turk J Haematol.Jun; 32(2): 172-4. Panel Bottega R, Marconi C et al. 2015. ACTN1-related thrombocytopenia: identification Pagel J, Beutel K et al. 2012. Distinct mutations in STXBP2 are associated of novel families for phenotypic characterization. Blood. Jan 29;125(5):748-50. with variable clinical presentations in patients with familial hemophagocytic lymphohistiocytosis type 5 (FHL5). Blood. 119:6016-6024. Cox K, Price V, Kahr WHA. 2011. Inherited platelet disorders: a clinical approach to diagnosis and management. Expert Rev Hematol. Aug;4(4):455-72. Paterson AD, Rommens JM, et al. 2010. Persons with Quebec platelet disorder have a tandem duplication of PLAU, the urokinase plasminogen activator gene. difficult to interpret, and typically require immediate testing on Faioni EM, Razzari C et al. 2014. Bleeding diathesis and gastro-duodenal ulcers Blood.115:1264-6. BloodCenter of Wisconsin offers a specifically in inherited cytosolic phospholipase-A2 alpha deficiency. Thromb Haemost. fresh patient platelets due to limited sample stability. Advances in Dec;112(6):1182-9. Rao AK. 2013. Inherited platelet function disorders: overview and disorders of designed Comprehensive Platelet Disorder granules, secretion, and signal transduction. Hematol Oncol Clin North Am. Jun; genetic testing through next-generation sequencing allows for Favier R, Raslova H. 2015. Progress in understanding the diagnosis and molecular 27(3):585-611. Panel (test code 4830) optimized for detection identification of underlying genetic defects and for distinguishing genetics of macrothrombocytopenias. Br J Haematol. Sep;170 (5):626-39. Rehm HL, Bale SJ et al. 2013. Working Group of the American College of Medical of germline variants in 43 genes known to inherited platelet disorder cases from immune thrombocytopenia. Freson K, Wijgaerts A, Van Geet C. 2014. Update on the causes of platelet disorders Genetics and Genomics Laboratory Quality Assurance Committee. ACMG clinical cause platelet function disorders and/or Accurate diagnosis provides information about the phenotype and functional consequences. John Wiley & Sons Ltd, Int. Jnl. Lab. Hem. 36, laboratory standards for next-generation sequencing. Genet Med.15:733-747. and prognosis, guides medical management decisions, assists 313–325. Richards S, Aziz N A et al.2015. Standards and guidelines for the interpretation of inherited thrombocytopenia. with the identification of affected family members, and allows for Gresele P. 2014. Diagnosis of inherited platelet function disorders: guidance for the sequence variants: a joint consensus recommendation of the American College accurate genetic recurrence risk assessment. SSC of the ISTH. J Thromb and Haemost. 13:314-322. of Medical Genetics and Genomics and the Association for Molecular Pathology. Gunay-Aygun M, Falik-Zaccai TC et al. 2011. NBEAL2 is mutated in Gray platelet Genet Med.17:405-424. Platelet function disorders and inherited thrombocytopenia are a Variants in several genes known to cause syndromic or non- Syndrome and is required for biogenesis of platelet alpha-granules. Nat Genet. Savoia A. 2016. Molecular basis of Inherited thrombocytopenias. Clin Genet heterogeneous group of disorders which may have overlapping syndromic platelet disorders may be inherited in an autosomal Aug; 43(8): 732–734. Feb;89(2):154-62. clinical phenotypes, generally differentiated by platelet counts. recessive, autosomal dominant or X-linked recessive manner. More Hinckley J, Di Paola J. 2014. Genetic basis of congenital platelet disorders. Savoia A. 2016. Molecular basis of Inherited thrombocytopenias: an update. Inherited thrombocytopenia disorders are typically characterized common and rare types of syndromic or non-syndromic platelet Hematology Am Soc Hematol Educ Program. Dec5;(1): 3337-42. Current Opinion in Hematology. Sep;23(5):486-492. by platelet counts less than 150,000/uL, but often can vary with disorders will be identified with this panel, including MYH9-related Israels SJ, El-Ekiaby M, Quiroga T, Mezzano D. 2010. Inherited disorders of platelet Kunishima Shinji S, Kobayashi R et al. 2009. Mutation of the β1-tubulin gene age, gender and ethnic background, while platelet function disorders, Glanzmann thrombasthenia, Bernard-Soulier syndrome, function and challenges to diagnosis of mucocutaneous bleeding. Haemophilia. associated with congenital macrothrombocytopenia affecting microtubule disorders are usually, but not always, associated with normal congenital amegakaryocytic thrombocytopenia, familial 16 (Suppl 5):152-9. assembly. Blood. 113:458-461. platelet counts and are caused by defects in platelet adhesion, platelet disorder with predisposition for acute myelogenous Johnson B, Lowe GC et al. 2016. Whole exome sequencing identifies genetic Songdej N, Rao AK. 2017. Inherited platelet dysfunction and hematopoietic glycoprotein expression, receptor function, signaling pathways, leukemia, ANKRD26-related thrombocytopenia, WAS-related variants in inherited thrombocytopenia with secondary qualitative function transcription factor mutations. Platelets Jan; 28(1):20-26. aggregation, cytoskeleton proteins, secretion, granular contents thrombocytopenia, Scott syndrome, gray platelet syndrome defects. Haematologica. 101(10):1170-1179. Stevenson WS, Rabbolini DJ et al. 2015. Paris-Trousseau thrombocytopenia is and abnormalities in procoagulant activity. Symptoms of platelet and others. Additional genes on this panel are associated with Kumar R, Kahr WHA. 2013. Congenital Thrombocytopenia: Clinical Manifestations, phenocopied by the autosomal recessive inheritance of a DNA-binding domain disorders may include purpura, petechiae, prolonged bleeding syndromes that have platelet disorders as a common finding Laboratory Abnormalities, and Molecular Defects of a Heterogeneous Group of mutation in FLI1. Blood. Oct 22;126(17); 2027-30. Conditions. Hematol Oncol Clin North Am. June; 27(3):465-94. from cuts, epistaxis, gum bleeding, excessive bleeding after among other non-hematologic features. Watson SP, Lowe M et al. 2013. Genotyping and phenotyping of platelet function surgery, hemoptysis, hematuria and menorrhagia in women. Kunicki TJ, Williams SA, Nugent DJ. 2012. Genetic variants that affect platelet disorders. J Thromb Haemost. 11(suppl.1):351-363. This panel evaluates for single nucleotide variants and small function. Current Opin Hematol.19:371-9. Severe platelet disorders can present in the newborn period, deletions and duplications, which are most commonly responsible Westbury SK, Mumford AD. 2016. Genomics of platelet disorders. Hemophilia. while mild thrombocytopenia may remain undiagnosed until Lentaigne C, Freson K et al. 2016. Inherited platelet disorders: toward DNA-based 22(Suppl.5): 20-24. for genetic disease. However, large deletions and duplications, diagnosis. Blood. 127(23):2814-2823. incidental detection of thrombocytopenia on routine blood also referred to as copy number variants (CNV), are a known cause Zhang My, Churpek JE et al. 2015. Germline ETV6 mutations in familial testing in adulthood. Some inherited platelet disorders have Manchev VT, Hilpert M et al. 2014. A new form of macrothrombocytopenia thrombocytopenia and hematologic malignancy. Nat Genet. Feb; 47(2):180-185. of genetic disorders, but can escape detection by next-generation induced by a germ-line mutation in the PRKACG gene. Blood. Oct 16; only hematologic manifestations, such as differences in platelet sequence analysis. Further testing with BloodCenter of Wisconsin 124(16):2554-63. Zhou Y, Zhang J. 2014. Arthrogryposis–renal dysfunction–cholestasis (ARC) size or distinctive granulocyte inclusions, while other syndromic syndrome: from molecular genetics to clinical features. Italian J Pediatrics. 40:77. custom designed, high density gene-focused array, aCGH Maclachlan A, Watson SP, Morgan NV. 2017. Inherited platelet disorders: Insight types present with additional non-hematologic manifestations. Deletion/Duplication Analysis, allows for the possible detection from platelet genomics using next-generation sequencing. Platelets. Jan ;28(1);14- Certain types of platelet disorders cause predisposition to acute 19. of large deletions and duplications within a single exon of a given myelogenous leukemia or myelodysplastic syndromes. gene, encompassing one or more exons, or affecting an entire Misdiagnosis of platelet disorders can result in inappropriate gene. This testing may be warranted when results of sequence therapies and inadequate surveillance for additional medical analysis do not fully explain a clinical phenotype, or when a complications, underscoring the importance of accurate suspected disorder is known to be caused by diagnosis. The diagnosis may be difficult to establish based solely on functional studies, especially in patients with milder disorders, as these assays are often technically challenging,