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    NRT.002 Saline Replacement

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    NRT.002 Saline Replacement

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    NRT.002-Saline-Replacement

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    NRT.002 Saline Replacement

    Uploaded by

    tailonglin1

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    of 3

    Ontario Regional Blood Coordinating Network

    Transfusion Technical Resource Manual

    Saline Replacement

    Approved By: Document No: NRT.002


    Date Issued: 2004/04/05 Category: Non-Routine Testing
    Date Revised: 2009/09/01; 2014/03/01 Page 1 of 3

    1.0. Principle

    To differentiate rouleaux from true agglutination.

    Rouleaux is a phenomenon that mimics agglutination. Red cells manifesting rouleaux


    appear microscopically like aggregates of stacked coins. It is caused by abnormalities in
    the patient’s plasma, not in the red cells. When the plasma is replaced with saline the
    rouleaux is dispersed but true agglutination remains.9.1

    Rouleaux is seen in diseases associated with abnormal serum protein concentrations.9.1

    2.0. Scope and Related Policies


    0.1 Normal saline replacement is usually done when all red cells tested at room
    temperature or 37°C show positive reactions with the patient’s plasma.

    0.2 Saline replacement is not done when all red cells tested by antiglobulin testing show
    positive reactions.

    3.0. Specimens
    EDTA anticoagulated whole blood

    4.0. Materials
    Equipment: Serological centrifuge
    Block for test tubes
    Microscope
    Vacuum aspiration (optional)

    Supplies: Test tubes – 10 x 75 mm, 12 x 75 mm


    Serological pipettes

    Reagents: Normal saline

    Ontario Regional Blood Coordinating Network NRT.002


    Transfusion Technical Resource Manual
    Page 1 of 3
    Saline Replacement
    5.0. Quality Control – N/A

    6.0. Procedure

    STEP ACTION
    6.1. Read tubes 6.1.1. Check agglutination for the characteristic “stacked coins”
    microscopically appearance
    6.2. Centrifuge the tubes 6.2.1. Centrifuge all tubes at 3400 rpm for 10-15 seconds.
    6.2.2. Using a pipette or vacuum aspiration equipment, remove all
    the supernatant from the centrifuged tubes leaving the cell
    button undisturbed
    6.3. Re-Suspend and Read 6.3.1. Add 2 drops of normal saline to each tube
    6.3.2. Re-suspend gently, read by initial reading method (i.e.,
    macroscopically or microscopically).
    6.4. Grade and record 6.4.1. See RT. 001 – Reading and Recording Hemagglutination
    results. Reactions.
    6.4.2. Indicate that the reading is after normal saline replacement
    by writing “SR” above the column where the results are
    being recorded.
    If Then
    entering results Footnote “reading performed after
    in a computer normal saline replacement.”
    6.5. Interpret the results 6.5.1. See 7.0 – Reporting.
    6.6. Perform a clerical 6.6.1. When the procedure is complete, perform a clerical check
    check. that:
     The patient name and identification number are
    identical on all specimens and the request form or
    computer screen
     The patient name is the same on all the test tubes and
    the request form or computer screen
     Verify that the test results have been interpreted
    correctly
    6.7. Initial 6.7.1. Initial or sign and record the completion time and date on
    the request form or in the computer.
    6.7.2. Verification of results must be recorded. See 7.0 Reporting.

    7.0. Reporting

    7.1 No agglutination after the normal saline replacement procedure usually indicates that
    rouleaux was present and has been dispersed by the normal saline. Report the
    reaction as negative.

    Ontario Regional Blood Coordinating Network NRT.002


    Standard Work Instruction Manual
    Page 2 of 3
    Saline Replacement
    7.2 Agglutination after the normal saline replacement procedure usually indicates true
    agglutination. Interpret the reaction as positive and continue with appropriate follow up
    (e.g., NRT.003 – ABO Group Problem Solving or antibody investigation if indicated).

    8.0. Procedural Notes – N/A

    9.0. References

    9.1 Roback JD, ed. AABBTechnical Manual, 17th ed. Bethesda, MD: American Association
    of Blood Banks, 2011: 488,901-902.

    9.2 Issitt PD, Anstee DJ. Applied Blood Group Serology, 4th ed. Durham, NC: Montgomery
    Scientific Publications, 1998: 1135.

    10.0 Revision History

    Revision Date Summary of Revision


    March 1, 2014  Revised name of manual
     Section 6.4.1- PA. 006 changed to
    RT.001
     Updated the list of references to include
    the most recent editions

    Ontario Regional Blood Coordinating Network NRT.002


    Standard Work Instruction Manual
    Page 3 of 3

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