Hemophilia B DNA Evaluation (Sendout)

General Information

Lab Name

Hemophilia B DNA Evaluation (Sendout)

Lab Code

RDHEMB

Epic Name

Hemophilia B DNA Evaluation (Sendout)

External Test Id

3250-10

Description

Test Information: Analysis of the Factor IX gene (Hemophilia B) is performed by direct sequence analysis of exons 1 – 8 and one common recurrent 3′ ut region genotype. The Genomics Testing Laboratory has had a high level of success in locating the mutation within these areas (Weinmann et al, BJ Haematol 100: 58, 1998).

    If the specific mutation cannot be identified, intragenic linkage is often useful to trace the affected allele through the family tree by following 4 polymorphic sites. Linkage studies also help identify the de novo origin of mutations in families with an isolated occurrence (eg maternal grandparents). Intragenic linkage may be useful in 80-90% of families where key family members (patient and both parents of potential carrier) are available.

      Additional Test Information: Sequencing is performed for exons 1-8 and the 3′ ut region. Confirmation is performed one of two ways. If the mutation creates/removes a restriction enzyme digestion site, the testing will be performed by RFLP. If a restriction enzyme digestion site is not present, testing will be done by sequencing the exon in which the mutation is located.

        Ordering Requirements:

        • Orders from Washington Center for Bleeding Disorders (SEAWBD), FHCC Hematology, and UW Medicine Prenatal Genetics are pre-approved. Orders from other locations require Laboratory Medicine Resident approval.
        • For orders not submitted via Epic or ISEAWBD interface, providers must complete a Bloodworks Eastlake Genomics Lab Requisition

        References

        1. Factor IX Gene (F9) Variant Database
        2. CDC Hemophilia Mutation Project: CHAMP & CHBMP
        3. Bloodworks Northwest Ordering Information: Bleeding Disorders

        Synonyms

        3250-10, F9 Genotyping, HemB Genotyping, Hemophilia B

        Components

        Code Name
        RHEMBS Hemophilia B DNA Evaluation Sample Type
        RHEMBR Hemophilia B DNA Evaluation Result
        RHEMBL Hemophilia B DNA Evaluation Performing Lab

        Interpretation

        Method

        Polymerase Chain Reaction (PCR) Based

        Reference Range

        See individual components

        Ref. Range Notes

        See report.

        Ordering & Collection

        Specimen Type

        Blood, Extracted DNA, Cultured Amniocytes

        Collection

        Whole Blood: 10 mL LAVENDER TOP EDTA tube

          Cultured Amniocytes: Two T-25 flasks of cells cultured to confluency.

          • Amniotic fluid is not accepted.
          • An EDTA whole blood sample from the mother of the fetus must accompany cultured amniocyte orders.

            Extracted DNA: >10 ug extracted DNA at a concentration of >25ng/ul

              Collection Timing:

              • Collect any time Monday through Thursday.
              • Samples collected on a Friday must be timed appropriately to ensure delivery at Bloodworks Eastlake Lab by the 1 p.m. cutoff.

                Samples sent on Fridays must be received at Bloodworks Northwest Genomics Lab by 1 p.m.

                Forms & Requisitions

                Approval Required

                Pre-approved for Washington Center for Bleeding Disorders (SEAWBD), FHCC Hematology, and UW Medicine Prenatal Genetics. Orders from other locations require LMR approval.

                Handling Instructions

                Outside Laboratories:

                • Store and transport whole blood at ambient temperature.
                • Expedite transport; samples must be received by Bloodworks NW within 72 hours of collection and before 1 p.m. on Fridays. Allow for one day of transport from UW-MT to Bloodworks Northwest.
                  • Unless the cells are cultured at UW-MT Cytogenetics and Genomics Lab, clients should arrange to send cultured amniocyte samples directly to Bloodworks NW due to the short stability window.
                • Submit a completed Bloodworks Northwest Eastlake Genomics Requisition along with the DLMP Requisition.* Leave the billing section blank to be completed by Sendouts staff.
                  • *Use client-specific requisition when available. To obtain a client-specific requisition, contact Client Support Services via email at: commserv@uw.edu.
                • WA Center for Bleeding Disorders only: A separate Bloodworks NW requisition is not required for orders submitted via interface.

                Stability (Whole Blood): Ambient (preferred): 72 hours; Refrigerated: 72 hours; Frozen: -20°C or -70/80°C. Avoid thawing/re-freezing.

                • Frozen EDTA tube(s) are accepted if samples are frozen the day of the blood draw, never thawed, and are shipped and received on dry ice. Contact Eastlake Bleeding Disorders at (206) 568-2184 with questions about sample acceptability.

                Quantity

                requested: 10 mL EDTA whole blood
                minimum: 3 mL EDTA whole blood

                Processing

                Result-at-Entry:

                • RHEMBS (Hemophilia B DNA Evaluation Sample Type): <Specimen type will file automatically for orders placed in Epic. For others, enter the appropriate specimen type code.>

                Specimen Type Codes:

                ETC Translation
                WB Whole Blood
                CULAMN Cultured Amniocytes
                DNAEXT Extracted DNA

                Processing:

                • Maintain whole blood and cultured amniocyte samples at ambient temperature.
                • Whole blood received frozen should be stored frozen.
                • If sample is collected on a Friday, notify Sendouts immediately upon sample receipt to ensure delivery to Bloodworks NW by the 1 p.m. cutoff.

                Sendouts:

                • Order Bloodworks NW Test: 3250-10
                • Submit a copy of the ISEAWBD eREQ or Epic requisition with the sample (ensure that all pages are printed to accommodate the order questions). For orders not placed in Epic or via the ISEAWBD interface, a separate Bloodworks Northwest Eastlake Genomics requisition is required.
                • Package sample(s) with the associated requisition in a biohazard bag (one patient per bag). Orders received with multiple
                  patient samples/requisitions in the same biozard bag will be rejected by Bloodworks NW.
                • Blood:
                  • Send blood orders as received with the UW DLMP or Delivery Express courier directly to the Bloodworks NW Eastlake location.
                  • Transport whole blood at ambient temperature. Whole blood received frozen should be stored frozen and transported on dry ice.
                  • Samples may be sent Monday through Friday only. If sent on a Friday, samples must be in lab at the Eastlake location by 1 p.m.
                  • Ambient or refrigerated whole blood samples must arrive at Bloodworks within 72 hours of collection.
                • Cultured amniocytes:
                  • Send cultured amniocytes same-day to the Bloodworks Central Genomics Lab (921 Terry Ave, Seattle WA 98104).
                  • Cultured amniocytes must arrive at Central Genomics within 48 hours of collection and by 1 p.m. on Fridays.

                Stability (Whole Blood): Ambient (preferred): 72 hours; Refrigerated: 72 hours; Frozen: -20°C or -70/80°C. Avoid thawing/re-freezing.

                • Frozen EDTA tube(s) are accepted if samples are frozen the day of the blood draw, never thawed, and are shipped and received on dry ice. Contact Eastlake Bleeding Disorders at (206) 568-2184 with questions about sample acceptability.

                Stability (Cultured Amniocytes): Samples must arrive within 48 hours to the Central Genomics location (amniocytes should not be sent to the Eastlake laboratory).

                Performance

                LIS Dept Code

                Sendouts (SO)

                Performing Location(s)

                Sendout Bloodworks Northwest Eastlake Genomics Laboratory
                206-568-2184

                1551 Eastlake Ave E
                Suite 100
                Seattle, WA 98102

                Also known as: Bloodworks Northwest Bleeding Disorders Laboratory

                Frequency

                Turn around Time: 5-10 business days from sample receipt at Bloodworks Eastlake Genomics.

                Available STAT?

                No

                Billing & Coding

                CPT codes

                81238

                LOINC

                18717-9

                Interfaced Order Code

                UOW5204