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Issue for the Identification of Antibodies

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Hello, fellow blood bankers.


Question for antibody identification of a patient with a previous history of transfusion!

What do you make of these results? and What are you going to do next?

We performed gel testing using Bio-Rad Liss/Coombs card and got some positive results in IAT.

Next, we performed tube testing with Ortho ID cells. We found strong reactivity at room temperature but some reactivity were present in IAT.

Autologous control is very weakly reactive at room temperature  (IgG and C3d DAT is negative). Do you think some reactivity in IAT (tube and gel) results from cold autoantibodies?

Or some IgG alloantibodies coexist with the cold autoantibodies? 

What do you make of these results? and what are you going to do next?


We have encountered such problems often. We really need your help.

Thank you. 




Edited by gebae
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From the second panel you posted, I think there are some specificity in the 37 and IAT phase, so my guessing it that there are not only cold auto that interfere with the reaction.

I will do an auto-absorption in 4 drgess first( If this patient has not received transfused red cells pack in three months), then run panel using tube IAT.

Just personal opinion, I am looking forward to learn from here.

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The possible antobodies are anti-E, V,K, Jsa and Lua, then I will pick some cells to test it and  antigen typing the patient to confirm it.( Jsa and Lua pos cells are quite rare, so it is hard to get them, luckily they will not cause rapid hemolysis after transfusion, as for anti-V, the clinical significance is unknown.)


Edited by yan xia
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I would treated the patient's own red cells with papain or ficin (whichever is used by the manufacturer to make their enzyme treated red cells), and then test them against autologous plasma.  My guess (and from this far away, it is just a guess) is that they will be positive.

I think that there is a "cold" reacting auto-antibody there.

I would also suggest performing an IAT panel in tubes, bringing the reactants to 37oC before mixing them, and using isotonic saline, rather than LISS as the red cell diluent, and then washing the tests in pre-warmed isotonic saline.  This should show if there are any underlying clinically significant alloantibodies.

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I would get more history first.

Transfused when? How much and/or how often? With what, rbcs, plasma, platelets, Immunoglobulin? Why? Diagnosis? Meds? Age of patient? Pregnancies if female? Previous antibody test results and methods utilized?  Any results of extended rbc testing previously done available? Any other lab results suggesting rbc destruction or decreased rbc survival of transfused red cells?

DAT negative but autocontrol positive? Could transfused red cells be present?  Early production of cold reactive auto or allo antibodies showing up?  As previously suggested try prewarming saline technique with no enhancement media, 45-60 min incubation with IgG reading only.  Could be cold and warm reactive auto or allo antibodies present.

Based on patient and testing histories should help with making a decision on what to do serologically.

It's a little troublesome if this type of reactivity is seen often in BB?  Unless there is some common issue with these patients, it may be something with regards to the actual methods or techniques used in the lab and not really a patient issue at all.

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Just a question - along with mrmic - What is the average daily temperature in your lab?  If this antibody is, at least partially, a "Cold" and you have a LOT of reactions like this, as you stated, it just may be too cold in your lab.  Try a strict Prewarm test, as suggested by Malcom and if that helps, consider reducing the amount of Room Temp exposure your average specimen encounters and see if you can cut the reactions down.  We have almost no RT exposure in our testing anymore (Immucor ECHO) but we used to have more extraneous reaction problems in the winter when our lab grew colder (think blue fingers!).  Just a suggestion.

Does anyone think this (second panel) may be an anti-P?  It is not showing the whole pattern, but I have seen several that didn't.

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