Shipping requirements References Ship on an ice pack or at room Andrikovics H, Klein I et al. 2003. Analysis of large structural changes of the factor temperature. Protect from freezing. VIII gene, involving intron 1 and 22, in severe hemophilia A. Haematologica. 88(7):778-84. Place the specimen and the requisition Bagnall RD, Giannelli F et al. 2006. Int22h-related inversions causing hemophilia into plastic bags and seal. Insert into A: a novel insight into their origin and a new more discriminant PCR test for their a Styrofoam container, seal and place detection. J Thromb Haemost.4(3)571-8 into a sturdy cardboard box, and Bagnall RD, Waseem N et al. 2002. Recurrent inversion breaking intron 1 of the tape securely. Ship the package in factor VIII gene is a frequent cause of severe hemophilia A. Blood. 99(1):168-74. Disorder compliance with your overnight carrier D P Bajaj SP, Thompson AR. 2006. Molecular and structural biology of factor IX. In: guidelines. Label with the following Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN, eds. Hemostasis and address: : Basic Principles and Clinical Practice. 5 ed. Philadelphia: Lippincott- Client Services/Diagnostic Laboratory Raven.131-150. BloodCenter of Wisconsin Bolton-Maggs PH et al. 2010. A study of variations in the reported A Panel 638 N. 18th St. prevalence around the world. Haemophilia.16:20–32. Milwaukee, WI 53233 Casini A, Blondon M, et al. 2015. Natural history of patients with congenital . Blood. Jan 15; 125(3): 553-561. de Moerloose P, Schved JF et al. 2016. Rare coagulation disorders: fibrinogen, factor BloodCenter of Wisconsin offers a specifically gene-focused array, aCGH Deletion/Duplication Analysis, allows Required forms VII and factor XIII. Haemophilia. Jul;22 Suppl 5:61-5. for the possible detection of large deletions and duplications Hua B, Fan L et al. 2009. Alpha 1- antitrypsin Pittsburgh in a family with designed Coagulation Disorder Panel (test Please complete all pages of the within a single exon of a given gene, encompassing one or more tendency. Haematologica. June; 94(6): 881-4. code 4815) optimized for the detection of requisition form. Clinical history exons, or affecting an entire gene. This testing may be warranted Lancellotti, S, Basso M, et al. 2013. Congenital prothrombin deficiency: an update. germline variants in 20 genes known to cause when results of sequence analysis do not fully explain a clinical (including patient’s ethnicity, clinical Semin. Thromb. Hemost. Sep;39(6):596-606. diagnosis, family history and relevant coagulation disorders. phenotype, or when a suspected disorder is known to be caused Lillicrap D. 2013. Molecular Testing for Disorders of Hemostasis. Int J Lab Hematol. by deletions or duplications. Please refer to the aCGH Deletion/ laboratory findings) is necessary for Jun; 35(3):290–296. optimal interpretation of genetic test Duplication Analysis test description for more information about Lillicrap D. 2013. : advances in pathogenetic understanding, Coagulation disorders, including hemophilia A and B, von specific genes included in this array. results and recommendations. Clinical diagnosis, and therapy. Blood. Nov 28;122 (23):3735-40. D and laboratory history can eitherP be Willebrand disease (VWD), and rare bleeding disorders (RBDs), Analysis of genes included in the Coagulation Disorder Panel may Neerman-Arbez M, de Moerloose P et al. 2016. Laboratory and Genetic are a heterogeneous group of inherited bleeding disorders with recorded on the requisition form or Investigation of Mutations Accounting for Congenital Fibrinogen Disorders. Semin also be ordered as a stand-alone single gene sequencing test clinical and laboratory reports can be Thromb Hemost.Jun;42(4):356-365. overlapping clinical phenotypes. Bleeding symptoms can include as dictated by the patient’s laboratory phenotype. Alternatively, included with the sample. Ng C, Motto DB et al. 2015. Diagnostic approach to von Willebrand disease. Blood. epistaxis, easy bruising, gingival bleeding, prolonged bleeding custom panels may be ordered if a patient’s history suggests a Mar 26;125(13):2029-2037. following an injury, surgery or dental extractions, gastrointestinal specific coagulation disorder with multiple causative genes, such Palla R, Peyvandi F et al. 2015. Rare bleeding disorders: diagnosis and treatment. or urinary bleeding, , , intracranial as fibrinogen deficiency, or if functional hemostasis results narrow CPT Codes/Billing/Turnaround time Blood. Mar 26; 125(13):2052-2061. bleeding and menorrhagia or postpartum bleeding in women. the diagnosis to specific phenotypes that can be due to different Symptoms can present at any age and range in severity: in mild Test Code: 4815 Peyvandi F, Kunicki T et al. 2013. Genetic sequence analysis of inherited bleeding underlying genetic conditions, as is seen with hemophilia A and diseases. Blood.1(22):3423-3431. cases, individuals remain asymptomatic until the event of a von Willebrand disease. Targeted familial variant testing can also CPT codes: 81240, 81241, 81332, 81355, 81400x5, 81401, 81405, Rehm HL, Bale SJ, et al. 2013. Working Group of the American College of Medical trauma or surgery, and in severe cases, patients may present with be performed on any gene in the panel when the specific genetic 81407, 81408, 81479 Genetics and Genomics Laboratory Quality Assurance Committee. ACMG clinical spontaneous life threatening hemorrhage or bleeding symptoms variant is known in a family. laboratory standards for next-generation sequencing. Genet Med.15:733-747. in the newborn period. Turnaround time: 21 days Additional types of inherited bleeding disorders associated Richards S, Aziz N et al. 2015. Standards and guidelines for the interpretation of Although results of functional hemostasis testing often guide The CPT codes provided are subject to change. CPT codes are sequence variants: a joint consensus recommendation of the American College with dysfunction are included in the Platelet Function provided only as guidance to assist clients with billing. of Medical Genetics and Genomics and the Association for Molecular Pathology. molecular testing for a specific inherited coagulation disorder, Disorder Panel. Both the Coagulation Disorder Panel and Platelet Genet Med.17(5):405-424. there are situations where functional tests are not definitive, Function Disorder Panel can be ordered together as part of the For additional information related to shipping, billing or pricing, are difficult to interpret, or may suggest two or more factor please contact, BloodCenter Client Services: (414) 937-6396 or Stonebraker JS, Bolton-Maggs PH et al. 2010. A study of variations in the reported Comprehensive Bleeding Disorder Panel. prevalence around the world. Haemophilia. Jan;16(1):20-32. deficiencies in a patient. For cases in which the laboratory 800-245-3117, Option 1, or [email protected]. phenotype is not fully consistent with clinical symptoms, Clinical Disorders combined factor deficiencies are suspected, or the specific Hemophilia A and B are X-linked disorders caused by pathogenic coagulation disorder is unclear, the Coagulation Disorder variants in the F8 and F9 genes. These disorders result in severe, Panel offers an efficient and cost-effective means of diagnostic moderate or mild factor VIII or factor IX deficiency primarily genetic evaluation. Accurate diagnosis provides information affecting males; however, female carriers can have varying about phenotype and prognosis, guides medical management degrees of factor VIII or factor IX deficiency with corresponding decisions, assists with the identification of affected family bleeding risk. The degree of plasma factor deficiency correlates members, and allows for accurate genetic recurrence risk with both the clinical severity of disease and genetic findings. assessment. This panel evaluates for single nucleotide variants and small deletions and duplications, which are most commonly responsible for genetic disease. However, large deletions and duplications, also referred to as copy number variants (CNV), are a known cause of genetic disorders, but can escape detection by next-generation sequence analysis. Further testing with the BloodCenter of Wisconsin custom designed, high density © Copyright 2017 BloodCenter of Wisconsin, Inc. , Part of Versiti. All rights reserved. 1 Severe hemophilia is characterized by plasma factor VIII or factor to quantitative defects of normal functioning VWF. Type 1C is Coagulation Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. IX levels of under 1 IU/dL. Moderate and mild hemophilia are associated with a significant increase of VWF clearance and characterized by factor VIII or factor IX levels of 1-5 IU/dL or 6-40 higher than normal response to DDAVP, but with a shortened FGA Pathogenic variants in FGA, FGB and FGG result in quantitative and/or qualitative 202400/ 134820 Autosomal Recessive IU/dL, respectively. Hemophilia B Leyden (caused by specific F9 half-life. Type 2 VWD is associated with qualitative defects of changes in the fibrinogen alpha, beta or gamma subunit chains respectively. Congenital afibrinogenemia: rare bleeding disorder with excessive bleeding often in the newborn variants) is associated with very low levels of functional factor IX VWF. Types 2A, 2B and 2M are characterized by mild-moderate period. Congenital hypofibrinogenimia results in decreased amounts of these subunit at birth, but hormonal changes cause factor IX levels to increase mucocutaneous bleeding. Types 2A and 2B are associated with chains and may lead to varying bleeding symptoms from mild-severe. gradually during puberty. Bleeding problems in adulthood are not loss of high molecular weight multimers, while in type 2M, Congenital dysfibrinogenemia and congenital hypodysfibrinogenemia may result in 616004/134820 Autosomal Dominant/ typically seen. normal multimers are present. In type 2B, bleeding symptoms, thromboembolic complications or both. Autosomal Recessive is often present. Type 2N is characterized by reduced factor VIII Genetic analysis is useful for identification of the underlying genetic FGB 202400/134830 Autosomal Recessive variant in males with severe, moderate or mild hemophilia for levels and associated with excessive bleeding following surgery. Type 2N may resemble a mild form of hemophilia A. Type 3 is 616004/134830 Autosomal Dominant/ determination of carrier status in the female individuals within their Autosomal Recessive families, as well as prenatal diagnosis for at-risk pregnancies. Genetic associated with severe bleeding due to essentially absent VWF results can also help predict risk of inhibitor development. When a and decreased factor VIII activity. Although most types of VWD FGG 202400/134850 Autosomal Recessive diagnosis of severe hemophilia A (congenital factor VIII deficiency are inherited in an autosomal dominant manner, some types of 616004/134850 Autosomal Dominant/ of <1%) is suspected or known, F8 Severe Hemophilia A Reflex 2A, type 2N, and type 3 are inherited in an autosomal recessive Autosomal Recessive Analysis is recommended. Approximately 50% of severe hemophilia manner. Please see individual test descriptions for more details GGCX Combined deficiency of dependent clotting factors-1(VKCFD1): mild to 277450/137167 Autosomal Recessive A cases are attributable to gene inversions in F8 introns 1 and 22. about VWD. severe bleeding symptoms including risk of episodes of perinatal/neonatal intracranial hemorrhage due to deficiency of factors II, VII, IX, and X. von Willebrand disease (VWD) is the most common inherited Rare bleeding disorders (RBDs) include inherited deficiencies Pseudoxanthoma elasticum-like disorder multiple coagulation factor deficiency: hyperlaxity 610842/137167 Autosomal Recessive bleeding condition in both males and females, affecting of coagulation proteins, fibrinogen, factors II, V, VII, X, XI, XIII, of the skin involving the entire body and deficiency of factors II, VII, IX and X. approximately 1% of the U.S. population. However, it is often combined coagulation factors involved in the intracellular transport of FV and FVIII, vitamin K dependent clotting factors, misdiagnosed, particularly among women. The disease is LMAN1 and factor VIII combined deficiency 1 (F5F8D1) and 2 (F5F8D2) are caused by 227300/601567 Autosomal Recessive characterized by either quantitative or qualitative defects of von plasminogen activator inhibitor, and alpha-2-plasmin inhibitor. pathogenic variants in LMAN1 and MCFD2 respectively: symptoms are usually mild and may Willebrand factor (VWF), a plasma protein that assists RBDs account for 3-5% of inherited coagulation disorders with include epistaxis, menorrhagia, and excessive bleeding during or after trauma. incidence varying between ethnicities. In most cases, RBDs are as they initially adhere at sites of vascular injury to begin MCFD2 613625/607788 Autosomal Recessive inherited in an autosomal recessive manner. the clotting process. The correct diagnosis of variant VWD is SERPINA1 Bleeding disorder of variable severity caused by specific variant SERPINA1 c.1145T>G 613490/107400 Autosomal Dominant essential to providing effective treatment. There are three types Refer to the table below for further information about each (p.Met358Arg) “Pittsburgh allele” resulting in an altered protein that inhibits , fXa of VWD, type 1, type 2 (2A, 2B, 2M and 2N), and type 3. Type 1 gene in the Coagulation Disorder Panel, including the clinical and protein C. Sequencing of the full coding region of SERPINA1 gene is not available. is typically associated with mild mucocutaneous bleeding due phenotype, OMIM numbers and inheritance pattern. SERPINE1 Plasminogen activator inhibitor-1 deficiency (PAI-1D): rare bleeding disorder associated 613329/173360 Autosomal Recessive/ with increased bleeding after trauma, injury or surgery due to increased fibrinolysis of fibrin Autosomal Dominant Coagulation Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. blood clots. Gene Clinical Phenotype Phenotype/Gene Inheritance SERPINF2 Alpha-2-plasmin inhibitor deficiency (APLID): severe increased susceptibility to bleeding. 262850/613168 Autosomal Recessive OMIM number VKORC1 Combined deficiency of vitamin K dependent clotting factors-2 (VKCFD2): mild to severe 607473/608547 Autosomal Recessive F2 Prothrombin deficiency: rare bleeding disorder with variable disease severity correlating 613679/176930 Autosomal Recessive bleeding symptoms including risk of perinatal/neonatal intracranial hemorrhage due to 1 with level of prothrombin (a vitamin K dependent glycoprotein) activity . deficiency of factors II, VII, IX, and X. 2 F5 Factor V deficiency: rare bleeding disorder with variable disease severity and age of onset . 227400/612309 Autosomal Recessive VWF von Willebrand disease (VWD): variable bleeding due to quantitative or qualitative defects F7 Factor VII deficiency: rare bleeding disorder with the most severe cases apparent in infancy. 227500/613878 Autosomal Recessive of von Willebrand factor (VWF). There are three types of VWD: Up to one-third of people with factor VII deficiency never have any bleeding problems. Type 1 VWD: accounts for more than 80% of cases; typically associated with mild 193400/613160 Autosomal Dominant Thrombotic episodes can occur in a minority of patients, usually with other thrombotic risk mucocutaneous bleeding due to reduction in quantity of normal functioning VWF. factors. Type VWD 1C: associated with significant increase of VWF clearance and higher than F8 Factor VIII deficiency (Hemophilia A): severe, moderate or mild bleeding disorder that 306700/300841 X-linked Recessive normal response to DDAVP, but has a shortened half-life. Autosomal Dominant primarily affects males. Female carriers may show varying degrees of factor VIII deficiency Type 2 VWD: qualitative defects of VWF, accounting for up to 10-15% of all cases of VWD, 613554/613160 (type 2B, 2M, and and related bleeding symptoms. with four primary subtypes: the majority of 2A); F9 Factor IX deficiency (Hemophilia B): severe, moderate or mild bleeding disorder primarily 306900/300746 X-linked Recessive • 2A: associated with mild-moderate mucocutaneous bleeding due to decreased VWF- Autosomal Recessive affecting males. Female carriers may show varying degrees of factor IX deficiency and dependent platelet adhesion with loss of high molecular weight multimers. (type 2N, some cases related bleeding symptoms. of type 2A) • 2B: associated with mild-moderate mucocutaneous bleeding with similar laboratory Hemophilia B Leyden (caused by specific F9 variants): associated with very low levels findings to type 2A but also increased VWF-platelet GPIb binding and often of functional factor IX at birth, but hormonal changes cause factor IX levels to increase thrombocytopenia. gradually during puberty. Bleeding problems in adulthood are not typically seen. • 2M: associated with mild-moderate mucocutaneous bleeding due to decreased VWF- F10 deficiency: rare bleeding disorder with variable disease severity associated with 227600/613872 Autosomal Recessive dependent platelet adhesion with normal distribution of multimers; a rare form of this reduced (type 1) or impaired (type II) factor X. type is characterized by decreased interaction with collagen. F11 Factor XI deficiency: typically presents with bleeding after trauma or surgery; homozygotes 612416/264900 Autosomal Recessive/ • 2N: characterized by a reduced factor VIII levels due to impaired ability of VWF to bind are more severely affected and there is variable bleeding problems in heterozygotes. Autosomal Dominant factor VIII; associated with excessive bleeding following surgery and can resemble a mild F13A1 Factor XIII deficiency: rare bleeding disorder with symptoms shortly after birth. Without 613225/134570 Autosomal Recessive form of hemophilia A. treatment, life-threatening intracranial hemorrhage may occur. Pathogenic variants in F13A1 and F13B severely reduce the amount or activity of the factor XIII A subunit or B Type 3 VWD: accounts for <5% of VWD and is associated with severe bleeding symptoms 277480/613160 Autosomal Recessive F13B subunit respectively. 613235/134580 Autosomal Recessive due to essentially absent VWF and decreased factor VIII activity.

1. The -related prothrombin variant F2 20210G>A in the 3’ 2. , the most common inherited form of thrombophilia will be untranslated region, which results in enhanced prothrombin production, will be detected with this assay. Individuals homozygous have an increased risk over 2 detected with this assay. heterozygotes. Severe hemophilia is characterized by plasma factor VIII or factor to quantitative defects of normal functioning VWF. Type 1C is Coagulation Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. IX levels of under 1 IU/dL. Moderate and mild hemophilia are associated with a significant increase of VWF clearance and characterized by factor VIII or factor IX levels of 1-5 IU/dL or 6-40 higher than normal response to DDAVP, but with a shortened FGA Pathogenic variants in FGA, FGB and FGG result in quantitative and/or qualitative 202400/ 134820 Autosomal Recessive IU/dL, respectively. Hemophilia B Leyden (caused by specific F9 half-life. Type 2 VWD is associated with qualitative defects of changes in the fibrinogen alpha, beta or gamma subunit chains respectively. Congenital afibrinogenemia: rare bleeding disorder with excessive bleeding often in the newborn variants) is associated with very low levels of functional factor IX VWF. Types 2A, 2B and 2M are characterized by mild-moderate period. Congenital hypofibrinogenimia results in decreased amounts of these subunit at birth, but hormonal changes cause factor IX levels to increase mucocutaneous bleeding. Types 2A and 2B are associated with chains and may lead to varying bleeding symptoms from mild-severe. gradually during puberty. Bleeding problems in adulthood are not loss of high molecular weight multimers, while in type 2M, Congenital dysfibrinogenemia and congenital hypodysfibrinogenemia may result in 616004/134820 Autosomal Dominant/ typically seen. normal multimers are present. In type 2B, thrombocytopenia bleeding symptoms, thromboembolic complications or both. Autosomal Recessive is often present. Type 2N is characterized by reduced factor VIII Genetic analysis is useful for identification of the underlying genetic FGB 202400/134830 Autosomal Recessive variant in males with severe, moderate or mild hemophilia for levels and associated with excessive bleeding following surgery. Type 2N may resemble a mild form of hemophilia A. Type 3 is 616004/134830 Autosomal Dominant/ determination of carrier status in the female individuals within their Autosomal Recessive families, as well as prenatal diagnosis for at-risk pregnancies. Genetic associated with severe bleeding due to essentially absent VWF results can also help predict risk of inhibitor development. When a and decreased factor VIII activity. Although most types of VWD FGG 202400/134850 Autosomal Recessive diagnosis of severe hemophilia A (congenital factor VIII deficiency are inherited in an autosomal dominant manner, some types of 616004/134850 Autosomal Dominant/ of <1%) is suspected or known, F8 Severe Hemophilia A Reflex 2A, type 2N, and type 3 are inherited in an autosomal recessive Autosomal Recessive Analysis is recommended. Approximately 50% of severe hemophilia manner. Please see individual test descriptions for more details GGCX Combined deficiency of vitamin K dependent clotting factors-1(VKCFD1): mild to 277450/137167 Autosomal Recessive A cases are attributable to gene inversions in F8 introns 1 and 22. about VWD. severe bleeding symptoms including risk of episodes of perinatal/neonatal intracranial hemorrhage due to deficiency of factors II, VII, IX, and X. von Willebrand disease (VWD) is the most common inherited Rare bleeding disorders (RBDs) include inherited deficiencies Pseudoxanthoma elasticum-like disorder multiple coagulation factor deficiency: hyperlaxity 610842/137167 Autosomal Recessive bleeding condition in both males and females, affecting of coagulation proteins, fibrinogen, factors II, V, VII, X, XI, XIII, of the skin involving the entire body and deficiency of factors II, VII, IX and X. approximately 1% of the U.S. population. However, it is often combined coagulation factors involved in the intracellular transport of FV and FVIII, vitamin K dependent clotting factors, misdiagnosed, particularly among women. The disease is LMAN1 Factor V and factor VIII combined deficiency 1 (F5F8D1) and 2 (F5F8D2) are caused by 227300/601567 Autosomal Recessive characterized by either quantitative or qualitative defects of von plasminogen activator inhibitor, and alpha-2-plasmin inhibitor. pathogenic variants in LMAN1 and MCFD2 respectively: symptoms are usually mild and may Willebrand factor (VWF), a plasma protein that assists platelets RBDs account for 3-5% of inherited coagulation disorders with include epistaxis, menorrhagia, and excessive bleeding during or after trauma. incidence varying between ethnicities. In most cases, RBDs are as they initially adhere at sites of vascular injury to begin MCFD2 613625/607788 Autosomal Recessive inherited in an autosomal recessive manner. the clotting process. The correct diagnosis of variant VWD is SERPINA1 Bleeding disorder of variable severity caused by specific variant SERPINA1 c.1145T>G 613490/107400 Autosomal Dominant essential to providing effective treatment. There are three types Refer to the table below for further information about each (p.Met358Arg) “Pittsburgh allele” resulting in an altered protein that inhibits thrombin, fXa of VWD, type 1, type 2 (2A, 2B, 2M and 2N), and type 3. Type 1 gene in the Coagulation Disorder Panel, including the clinical and protein C. Sequencing of the full coding region of SERPINA1 gene is not available. is typically associated with mild mucocutaneous bleeding due phenotype, OMIM numbers and inheritance pattern. SERPINE1 Plasminogen activator inhibitor-1 deficiency (PAI-1D): rare bleeding disorder associated 613329/173360 Autosomal Recessive/ with increased bleeding after trauma, injury or surgery due to increased fibrinolysis of fibrin Autosomal Dominant Coagulation Disorder Panel: gene, clinical phenotype, OMIM number and inheritance pattern. blood clots. Gene Clinical Phenotype Phenotype/Gene Inheritance SERPINF2 Alpha-2-plasmin inhibitor deficiency (APLID): severe increased susceptibility to bleeding. 262850/613168 Autosomal Recessive OMIM number VKORC1 Combined deficiency of vitamin K dependent clotting factors-2 (VKCFD2): mild to severe 607473/608547 Autosomal Recessive F2 Prothrombin deficiency: rare bleeding disorder with variable disease severity correlating 613679/176930 Autosomal Recessive bleeding symptoms including risk of perinatal/neonatal intracranial hemorrhage due to 1 with level of prothrombin (a vitamin K dependent glycoprotein) activity . deficiency of factors II, VII, IX, and X. 2 F5 Factor V deficiency: rare bleeding disorder with variable disease severity and age of onset . 227400/612309 Autosomal Recessive VWF von Willebrand disease (VWD): variable bleeding due to quantitative or qualitative defects F7 Factor VII deficiency: rare bleeding disorder with the most severe cases apparent in infancy. 227500/613878 Autosomal Recessive of von Willebrand factor (VWF). There are three types of VWD: Up to one-third of people with factor VII deficiency never have any bleeding problems. Type 1 VWD: accounts for more than 80% of cases; typically associated with mild 193400/613160 Autosomal Dominant Thrombotic episodes can occur in a minority of patients, usually with other thrombotic risk mucocutaneous bleeding due to reduction in quantity of normal functioning VWF. factors. Type VWD 1C: associated with significant increase of VWF clearance and higher than F8 Factor VIII deficiency (Hemophilia A): severe, moderate or mild bleeding disorder that 306700/300841 X-linked Recessive normal response to DDAVP, but has a shortened half-life. Autosomal Dominant primarily affects males. Female carriers may show varying degrees of factor VIII deficiency Type 2 VWD: qualitative defects of VWF, accounting for up to 10-15% of all cases of VWD, 613554/613160 (type 2B, 2M, and and related bleeding symptoms. with four primary subtypes: the majority of 2A); F9 Factor IX deficiency (Hemophilia B): severe, moderate or mild bleeding disorder primarily 306900/300746 X-linked Recessive • 2A: associated with mild-moderate mucocutaneous bleeding due to decreased VWF- Autosomal Recessive affecting males. Female carriers may show varying degrees of factor IX deficiency and dependent platelet adhesion with loss of high molecular weight multimers. (type 2N, some cases related bleeding symptoms. of type 2A) • 2B: associated with mild-moderate mucocutaneous bleeding with similar laboratory Hemophilia B Leyden (caused by specific F9 variants): associated with very low levels findings to type 2A but also increased VWF-platelet GPIb binding and often of functional factor IX at birth, but hormonal changes cause factor IX levels to increase thrombocytopenia. gradually during puberty. Bleeding problems in adulthood are not typically seen. • 2M: associated with mild-moderate mucocutaneous bleeding due to decreased VWF- F10 Factor X deficiency: rare bleeding disorder with variable disease severity associated with 227600/613872 Autosomal Recessive dependent platelet adhesion with normal distribution of multimers; a rare form of this reduced (type 1) or impaired (type II) factor X. type is characterized by decreased interaction with collagen. F11 Factor XI deficiency: typically presents with bleeding after trauma or surgery; homozygotes 612416/264900 Autosomal Recessive/ • 2N: characterized by a reduced factor VIII levels due to impaired ability of VWF to bind are more severely affected and there is variable bleeding problems in heterozygotes. Autosomal Dominant factor VIII; associated with excessive bleeding following surgery and can resemble a mild F13A1 Factor XIII deficiency: rare bleeding disorder with symptoms shortly after birth. Without 613225/134570 Autosomal Recessive form of hemophilia A. treatment, life-threatening intracranial hemorrhage may occur. Pathogenic variants in F13A1 and F13B severely reduce the amount or activity of the factor XIII A subunit or B Type 3 VWD: accounts for <5% of VWD and is associated with severe bleeding symptoms 277480/613160 Autosomal Recessive F13B subunit respectively. 613235/134580 Autosomal Recessive due to essentially absent VWF and decreased factor VIII activity.

1. The thrombophilia-related prothrombin variant F2 20210G>A in the 3’ 2. Factor V Leiden, the most common inherited form of thrombophilia will be untranslated region, which results in enhanced prothrombin production, will be detected with this assay. Individuals homozygous have an increased risk over 3 detected with this assay. heterozygotes. Indications for testing Assay sensitivity and limitations Coagulation Disorder Panel: The analytical sensitivity of this test is >99% for single nucleotide • Clarification and/or confirmation of diagnosis in a patient with changes and insertions and deletions of less than 20 bp. This clinical and laboratory findings of a bleeding disorder when assay does not detect large deletions or duplications (>20 bp), patient’s history suggests multiple coagulation disorders or deletions, duplications or variants that are outside the regions sequenced. To order analysis of copy number variation at the • Identification of carriers with family history of unspecified exon or gene level, please refer to the aCGH Deletion/Duplication bleeding disorders to provide accurate reproductive risk Analysis test or contact Client Services before placing your order. assessment and genetic counseling Please see hemophilia A, hemophilia B and VWD individual test Single gene sequencing or custom gene panel: descriptions for additional information about analytical and clinical sensitivity for these disorders. • Analysis of genes included in the Coagulation Disorder Panel may also be ordered as a stand-alone single gene sequencing test or custom panel (2-10 genes) as dictated by the patient’s Reporting of results clinical and laboratory phenotype While this assay is designed to detect germline genetic variants Targeted familial variant analysis: associated with coagulation disorders, variants unrelated to the • Targeted variant analysis for clinical diagnosis, carrier indication for testing, but with other clinical and/or reproductive identification or prenatal diagnosis can also be performed on implications, may also be detected. A comprehensive database of any gene in the panel when the pathogenic variant(s) is known gene-phenotype relationships listed by gene name can be found in the family (test code: 4970) at http://www.omim.org. For clinical questions about laboratory tests and test utilization Results are classified and reported in accordance with ACMG support, contact BloodCenter Client Services: (414) 937-6396 or next-generation sequencing standards. Variants predicted to 800-245-3117, Option 1, to be directed to our genetic counselors be pathogenic, likely pathogenic, and of uncertain significance and clinical support team. will be reported; variants classified as likely benign or benign are typically not reported but such data are available upon request. Test method Sequence variants are described using standard Human Genome Variation Society (HGVS) nomenclature (http://hgvs.org). This next-generation sequencing assay analyzes 19 genes, spanning the full coding regions plus a minimum 30bp of non-coding DNA including intron-exon junctions, in addition Specimen requirements to 67 bp upstream of F9 exon 1 to cover F9 Leyden variants, the Parental/Patient/Pediatric: 3-5 mL Whole Blood (EDTA tube, 3’ UTR of F2 and the 5’ UTR of F8 and VWF, as well as targeted lavender top), 2-5 mL Bone Marrow (EDTA tube, lavender top), 3-4 sequencing of SERPINA1 c.1145T. These targeted regions Buccal Swabs or ≥1ug of DNA at ≥50ng/uL of High Quality DNA. are captured by hybridization, amplified and sequenced by massively parallel sequencing. Regions will have a minimum Fetal: 7-15 mL Amniotic fluid, 5-10 mg Chorionic villi; back up coverage of 50x and those regions with less than 50 sequencing culture of amniocytes or chorionic villi is highly recommended. reads or low quality coverage are supplemented with Sanger Cultured: Two T25 flasks cultured amniocytes or cultured chorionic sequencing. All regions are covered by bi-directional analysis. villi (2x106 minimum). Maternal Blood sample of 3-5 mL Whole Variants are identified by a customized bioinformatics pipeline, Blood (EDTA tube, lavender top) is requested for all prenatal analyzed and comprehensively interpreted by our team of samples for maternal cell contamination studies. directors, scientists, and genetic counselors. All reported variants, If questions, please contact the laboratory to discuss sample including pathogenic, likely pathogenic, and variants of uncertain requirements. 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.

4 Shipping requirements Andrikovics H, Klein I et al. 2003. Analysis of large structural changes of the factor VIII gene, involving intron 1 and 22, in severe hemophilia A. Haematologica. Ship on an ice pack or at room 88(7):778-84. temperature. Protect from freezing. Bagnall RD, Giannelli F et al. 2006. Int22h-related inversions causing hemophilia Place the specimen and the requisition A: a novel insight into their origin and a new more discriminant PCR test for their Coagulation into plastic bags and seal. Insert into detection. J Thromb Haemost.4(3)571-8 a Styrofoam container, seal and place Bagnall RD, Waseem N et al. 2002. Recurrent inversion breaking intron 1 of the into a sturdy cardboard box, and factor VIII gene is a frequent cause of severe hemophilia A. Blood. 99(1):168-74. tape securely. Ship the package in Bajaj SP, Thompson AR. 2006. Molecular and structural biology of factor IX. In: Colman RW, Hirsh J, Marder VJ, Clowes AW, George JN, eds. Hemostasis and Disorder compliance with your overnight carrier D P Thrombosis: Basic Principles and Clinical Practice. 5 ed. Philadelphia: Lippincott- guidelines. Label with the following Raven.131-150. address: Bolton-Maggs PH et al. 2010. A study of variations in the reported haemophilia A Client Services/Diagnostic Laboratory prevalence around the world. Haemophilia.16:20–32. BloodCenter of Wisconsin Panel Casini A, Blondon M, et al. 2015. Natural history of patients with congenital 638 N. 18th St. dysfibrinogenemia. Blood. Jan 15; 125(3): 553-561. Milwaukee, WI 53233 de Moerloose P, Schved JF et al. 2016. Rare coagulation disorders: fibrinogen, factor VII and factor XIII. Haemophilia. Jul;22 Suppl 5:61-5. BloodCenter of Wisconsin offers a specifically gene-focused array, aCGH Deletion/Duplication Analysis, allows Required forms Hua B, Fan L et al. 2009. Alpha 1- antitrypsin Pittsburgh in a family with bleeding tendency. Haematologica. June; 94(6): 881-4. designed Coagulation Disorder Panel (test for the possible detection of large deletions and duplications within a single exon of a given gene, encompassing one or more Please complete all pages of the Lancellotti, S, Basso M, et al. 2013. Congenital prothrombin deficiency: an update. code 4815) optimized for the detection of requisition form. Clinical history Semin. Thromb. Hemost. Sep;39(6):596-606. exons, or affecting an entire gene. This testing may be warranted (including patient’s ethnicity, clinical Lillicrap D. 2013. Molecular Testing for Disorders of Hemostasis. Int J Lab Hematol. germline variants in 20 genes known to cause when results of sequence analysis do not fully explain a clinical diagnosis, family history and relevant Jun; 35(3):290–296. coagulation disorders. phenotype, or when a suspected disorder is known to be caused laboratory findings) is necessary for Lillicrap D. 2013. von Willebrand disease: advances in pathogenetic understanding, by deletions or duplications. Please refer to the aCGH Deletion/ optimal interpretation of genetic test diagnosis, and therapy. Blood. Nov 28;122 (23):3735-40. Duplication Analysis test description for more information about results and recommendations. Clinical Neerman-Arbez M, de Moerloose P et al. 2016. Laboratory and Genetic Coagulation disorders, including hemophilia A and B, von specific genes included in this array. and laboratory history can either be Investigation of Mutations Accounting for Congenital Fibrinogen Disorders. Semin Willebrand disease (VWD), and rare bleeding disorders (RBDs), Analysis of genes included in the Coagulation Disorder Panel may D P Thromb Hemost.Jun;42(4):356-365. recorded on the requisition form or are a heterogeneous group of inherited bleeding disorders with also be ordered as a stand-alone single gene sequencing test clinical and laboratory reports can be Ng C, Motto DB et al. 2015. Diagnostic approach to von Willebrand disease. Blood. overlapping clinical phenotypes. Bleeding symptoms can include as dictated by the patient’s laboratory phenotype. Alternatively, Mar 26;125(13):2029-2037. included with the sample. epistaxis, easy bruising, gingival bleeding, prolonged bleeding custom panels may be ordered if a patient’s history suggests a Palla R, Peyvandi F et al. 2015. Rare bleeding disorders: diagnosis and treatment. following an injury, surgery or dental extractions, gastrointestinal specific coagulation disorder with multiple causative genes, such Blood. Mar 26; 125(13):2052-2061. or urinary bleeding, hematomas, hemoptysis, intracranial as fibrinogen deficiency, or if functional hemostasis results narrow CPT Codes/Billing/Turnaround time Peyvandi F, Kunicki T et al. 2013. Genetic sequence analysis of inherited bleeding bleeding and menorrhagia or postpartum bleeding in women. the diagnosis to specific phenotypes that can be due to different diseases. Blood.1(22):3423-3431. Symptoms can present at any age and range in severity: in mild Test Code: 4815 underlying genetic conditions, as is seen with hemophilia A and Rehm HL, Bale SJ, et al. 2013. Working Group of the American College of Medical cases, individuals remain asymptomatic until the event of a von Willebrand disease. Targeted familial variant testing can also CPT codes: 81405, 81407, 81408, 81479 Genetics and Genomics Laboratory Quality Assurance Committee. ACMG clinical trauma or surgery, and in severe cases, patients may present with laboratory standards for next-generation sequencing. Genet Med.15:733-747. be performed on any gene in the panel when the specific genetic Turnaround time: 21 days spontaneous life threatening hemorrhage or bleeding symptoms variant is known in a family. Richards S, Aziz N et al. 2015. Standards and guidelines for the interpretation of in the newborn period. The CPT codes provided are subject to change. CPT codes are sequence variants: a joint consensus recommendation of the American College Additional types of inherited bleeding disorders associated provided only as guidance to assist clients with billing. of Medical Genetics and Genomics and the Association for Molecular Pathology. Although results of functional hemostasis testing often guide with platelet dysfunction are included in the Platelet Function Genet Med.17(5):405-424. molecular testing for a specific inherited coagulation disorder, For additional information related to shipping, billing or pricing, Disorder Panel. Both the Coagulation Disorder Panel and Platelet Stonebraker JS, Bolton-Maggs PH et al. 2010. A study of variations in the reported there are situations where functional tests are not definitive, Function Disorder Panel can be ordered together as part of the please contact, BloodCenter Client Services: (414) 937-6396 or haemophilia A prevalence around the world. Haemophilia. Jan;16(1):20-32. are difficult to interpret, or may suggest two or more factor Comprehensive Bleeding Disorder Panel. 800-245-3117, Option 1, or [email protected]. deficiencies in a patient. For cases in which the laboratory References phenotype is not fully consistent with clinical symptoms, Clinical Disorders combined factor deficiencies are suspected, or the specific Hemophilia A and B are X-linked disorders caused by pathogenic coagulation disorder is unclear, the Coagulation Disorder variants in the F8 and F9 genes. These disorders result in severe, Panel offers an efficient and cost-effective means of diagnostic moderate or mild factor VIII or factor IX deficiency primarily genetic evaluation. Accurate diagnosis provides information affecting males; however, female carriers can have varying about phenotype and prognosis, guides medical management degrees of factor VIII or factor IX deficiency with corresponding decisions, assists with the identification of affected family bleeding risk. The degree of plasma factor deficiency correlates members, and allows for accurate genetic recurrence risk with both the clinical severity of disease and genetic findings. assessment. This panel evaluates for single nucleotide variants and small deletions and duplications, which are most commonly responsible for genetic disease. However, large deletions and duplications, also referred to as copy number variants (CNV), are a known cause of genetic disorders, but can escape detection by next-generation sequence analysis. Further testing with the BloodCenter of Wisconsin custom designed, high density © Copyright 2017 BloodCenter of Wisconsin, Inc. , Part of Versiti. All rights reserved.