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Seeing Beyond Limits

2020 Webinar Schedule

Link to register:  https://immucor.webinato.com/register

Seeing Beyond Limits

2020 “Mini” WebinarsShort,10-minute educational videos with an industry leader.

Link to register:  https://immucor.webinato.com/register

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Continuing Education

• PACE, Florida and California DHS

• 1.0 Contact Hours

• Each attendee must register to receive CE at:https://www.surveymonkey.com/r/ImmunohematologyCaseStudies

• Registration deadline is July 3, 2020

• Certificates will be sent via email only to those who have registered July 10, 2020

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• Course content is for information and illustration purposes only. Immucor makes no representation or warranties about the accuracy or reliability of the information presented, and this information is not to be used for clinical or maintenance evaluations.

• The opinions contained in this presentation are those of the presenter and do not necessarily reflect those of Immucor.

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Cami Melland MLS(ASCP)SBBImmunohematology Reference Lab Manager

Q4 2019

Capture R to the Rescue!

Vitalant exists to help people realize their life-transforming potential by offering convenient blood donation opportunities and sharing our expertise in transfusion medicine. Founded in 1943, Vitalant is one of the nation’s oldest and largest nonprofit transfusion medicine organizations. We comprise a network of nearly a dozen community blood centers, each with its own rich history and legacy. Vitalant is an internationally known and respected leader that continues to push the boundaries to advance the experience, practice and application of transfusion medicine.

https://vitalant.org/

Immunohematology Reference Laboratories (IRL)

http://laboratories.vitalant.org/

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• 15 staff members • Provides services 24/7 for Colorado, Wyoming and South Dakota hospitals primarily• Completes over 5500 patient workups, sends approximately 750 special platelets and 5000 antigen negative RBCs each year

Denver IRL

Patient

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Pre-transfusion testing for hip surgery on a 77 year old female reveals pan-reactivity in gel. A cold screen provided by the facility was negative. The sample was sent to the Denver IRL for testing. The patient had a history of a warm autoantibody with no underlying alloantibodies 8 years prior.

Initial Testing

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• Weak, mixed field-like, refractile agglutination exhibited with 60% of the red cells tested using LISS enhancement at the AHG phase.

• The autocontrol was NOT positive indicating that the previously identified warm autoantibody is no longer showing.

• What’s next?

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Anti-Sda Investigation

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• Weak, mixed field-like, refractile reactivity at AHG is an unusual finding and can often can be attributed to the presence of anti-Sda.

• Anti-Sda is directed against an antigen found in tissues, body fluids and on red cells of more than 91% of adults.

• The antibody has been shown to have little clinical significance.

• Patients with anti-Sda do not require Sd(a-) blood for transfusion. Often present with a cold autoantibody and/or during pregnancy.

Urine Preparation

https://en.wikipedia.org/wiki/Dialysis_(biochemistry)17

•Urine neutralization has been shown to

neutralize anti-Sda.

•The AABB Technical Manual provides

instruction on how to prepare the urine.

•Pool urine from at least 6 individuals of known

Sda type.

•Boil it for 10 min and then cool.

•Dialyze it against phosphate buffered saline

(PBS) at 4C for 48 hours.

•Centrifuge and store at -20C.

Urine Neutralization

• Mix equal volumes of thawed urine and patient plasma.

• Prepare a control tube containing equal volumes of patient plasma with PBS.

• Incubate at room temp for 30 min.

• Test the control and urine plasma against red cells.

• If the control is positive and the urine plasma is negative anti-Sda was neutralized.

• Red Cell Alloantibodies cannot be ruled out using neutralized plasma! Too dilute.

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Urine Neutralization Results

• What do these results indicate?

• Partial neutralization

• Presence of anti-Sda and another antibody??

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Weak Anti-Sda

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• Anti-Sda exists at differing strengths:

• Weak: Alloantibodies can be ruled out with PEG and/or LISS testing with reagent red cells.

• ABO group compatible units can be provided for transfusion. The crossmatch results may or may not be compatible.

Strong Anti-Sda

• Strong: Alloantibodies cannot be ruled out using PEG and/or LISS enhancement

• Patient phenotype obtained if possible

• Phenotypically matched, antigen typed red cells provided for transfusion

• Crossmatch will be reported as incompatible

• If the patient has been transfused recently, molecular testing suggested

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Manual Capture R

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• Solid phase added to the Denver Immunohematology Reference Lab (IRL) tool box in 2016

• 62% of clients used solid phase testing and wanted the IRL to be able to replicate reactivity

• Long expiration dates

• Ability to create your own monolayer with any red cells

• Add saline and red cells to well

• Spin and wash-VOILA!

• Test

• Add LISS and patient plasma

• Incubate at 37C and then wash

• Add indicator cells and spin

Troubles Detected Using Tube Methodology

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• Cold autoantibodies (IgM) carrying through to the antiglobulin (AHG) phase of testing

• Antibodies previously known an “High-Titer Low Avidity” (HTLA-like) reacting with nearly 100% of cells at AHG

• Anti-Sda

• Antibodies to reagent red cell diluent

• These examples are generally notclinically significant and not picked up by manual solid phase

Capture R to the Rescue!

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The End

• Now we can rest easy knowing that the reactivity is not due to an underlying alloantibody.

• A full patient phenotype is not required.

• Antigen matched units is not required for transfusion.

• Money, time and special red cell products saved.

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References

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Technical Manual. Current ed. Bethesda, MD: AABB, 2011: 433, 477

Issitt PD, Issitt CH. Applied Blood Group Serology. 4th ed. Montgomery Scientific Publications Durham, NC. 1998: 814-818.

Mollison PL. Blood Transfusion in Clinical Medicine. 8th ed. Oxford: Blackwell Scientific. 1987: 265.

Harmening, D. 2012. Modern Blood Banking and Transfusion Practices. 6 th Edition. Philadelphia: F.A. Davis Company

Immunohematology Methods and Procedures. 1st Edition, 1993. Rockville: American Red Cross National Reference Laboratory.

Issitt, Peter D. and Anstee, David J. 1998. Applied Blood Group Serology. Fourth Edition. Durham: Montgomery Scientific Publications.

Judd, JW. 2008. Methods in Immunohematology. 3rd Edition.

Wallace ME, Green TS, eds. Selection of procedures for problem solving. Arlington, VA: AABB, 1983: 120-1.

Questions?

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vitalant.org

Thank you

Anti‐Jka in Capture

Louanne Shannon BS, MLS, MT(ASCP)SBBTransfusion Service Section Chief

CASE• Patient History:

• Anti‐D 

• Cold antibody of unknown specificity 

• Ready Screen

• Cells I and II positive

• Cell III negative  

• Technologist :

• New Generalist on duty in Blood Bank

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Previous Anti-D & Cold Antibody

Ready ID on the ECHO

Rule Outs Needed

When the tech could not rule out on the ECHO:

• Anti‐Fya, 

• Anti‐Jka ,or 

• Anti‐Lea

A tube panel was performed using PeG.  

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Negative Tube PanelPositive ECHO Crossmatches

New Technologist

Suspected antibody towards stroma

Crossmatched with PeG= Crossmatches compatible with PeG

Ready ID on the ECHO

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Supervisory ReviewSuspect:

Anti-Jka on ECHO ReadyID Panel

Ruled Out:

Anti- Fya : patient sample (Fya+)

Anti-Lea: tube panel

Confirmed:

Anti- Jka : Patient sample (Jka-)

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Continuing Education

• PACE, Florida and California DHS

• 1.0 Contact Hours

• Each attendee must register to receive CE at:https://www.surveymonkey.com/r/ImmunohematologyCaseStudies

• Registration deadline is July 3, 2020

• Certificates will be sent via email only to those who have registered July 10, 2020

39All Content © Immucor

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41All Content © Immucor

Thank you!