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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.

Cami Melland MLS(ASCP)SBB

Immunohematology 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/

• 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

14

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

15

• 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?

Anti-Sda Investigation

16

• 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.

18

Urine Neutralization Results

• What do these results indicate?

• Partial neutralization

• Presence of anti-Sda and another antibody??

19

Weak Anti-Sda

20

• 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

21

Manual Capture R

22

• 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

23

• 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 not

clinically significant and not picked up by

manual solid phase

Capture R to the Rescue!

24

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.

25

References

26

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?

27

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

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.

Negative Tube Panel

Positive ECHO Crossmatches

New Technologist

Suspected antibody towards stroma

Crossmatched with PeG

= Crossmatches compatible with PeG

Ready ID on the ECHO

Supervisory Review

Suspect:

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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Thank you!