BloodBankBlake

We use regular blood bank saline or whatever suspension the reagent RBCs are in (as long as it isn't a LISS suspension). The procedure is just doing a screen/panel/xm don't add enhancement media, and incubate for 30min@ 37C

From my manager's "old school" approach that if a clinically significant antibody is present with a warm auto, it will react alone without enhancement media due to its supposed "high" titer. Similar to slsmith, we incorporate this approach when we have panagglutination in our more sensitive methodologies and are trying to "look through" these reactions. It's never our sole method or what we use to report, but just another arrow in the quiver of techniques. 

and then give the rest of us a book report!  

I think you will find that dichloro-diphenyl-trichloroethane will destroy a lot more than just the Kell Blood Group System antigens!!!!!!!!!!!  

If your patient is Kell Negative (Ko) you have real problems.  If your patient is K Negative, you, and your patient, have more chance!

Some good materials here as well => https://www.bbguy.org/2016/06/17/want-g-wiz/

In particular, guinea pig urine.  How you get this is another matter (even if you have a guinea pig) because 1) they keep moving and 2) they won't urinate to order!!!!!!!!!!!

Usually Sda agglutination has a distinctive appearance.  You can also neutralize it with urine (as I recall).  Just pool a bunch of urines and you are bound to have an effective neutralizing soln. (better if you learn to recognize its appearance)

I am enormously honoured to announce that I am going to be awarded the Gold Medal of the British Blood Transfusion Society at their Annual Scientific Meeting in Brighton this year.  It is awarded to an individual for their exceptional and long standing services to the Society and to the practice of blood transfusion in the UK.  Sorry if this sounds egocentric, but I am very excited.

This is me going back to the early 1970s, when I was first working at the International Blood Group Reference Laboratory (IBGRL) when it was still in London, and so my memory might not be 100% reliable, but I'll have a go.
Obviously, in those days we used to use polyclonal anti-A or anti-B that was derived from human donors, rather than monoclonal antibodies (essentially, there were no such things in those days).  We did know enough, however, not to use anti-A,B from group O donors, because of the chance of cross-reaction.
We incubated the red cells against the antibodies at 4oC for about an hour, and then washed them at least six times in normal saline (we didn't even use buffered saline in those days).  The last wash was kept as a negative control.
Next comes the clever bit.  We used heat elution at 56oC, as did many people, but the really clever bit was the way we kept the supernatant at 56oC during the centrifugation stage, so that the antibodies would not go back onto the antigens as the tests cooled down.  The Director of the IBGRL at the time was the late, GREAT Dr Kenneth Goldsmith (I use the term "great" advisedly).  He built a wooden box that contained the centrifuge, but also had a common light bulb in it that heated the entire contraption when turned on (we had to turn it on a good half an hour before we used it, to allow it all to come to temperature), but the whole contained a thermostat, so that the temperature as close to 56oC, so that a higher temperature did not denature the antibody, and a lower temperature did not allow the antibody to go back on to the antigens (he really was an amazing person - a doctor who was also an expert scientist).
The eluate and the last wash were then tested against A1, B and O red cells at room temperature (23oC incubator), which, of course, all acted as internal controls.
Later, we realised that the Lui elution technique (using melting ice) was a more efficient technique for eluting ABO antibodies, but everything else (barring the heated centrifuge) was the same.
Nowadays, we hardly ever bother.  It is very time consuming for virtually no return.  If the subject is a patient, we would transfused group O red cells, and if the subject is a donor, it is cheaper and simpler to exclude them, as long as they are counselled, to ensure that they do not feel "stigmatised".

I just answered this question.


My Score PASS  

Firstly, and if possible, we would use an IgM monoclonal agglutinating antibody.
Secondly, we can elute off the auto-antibody using mild treatment with DTT or ZZAP, leaving the red cells relatively intact (relatively, because some antigens are denatured by this, while some others are considerably weakened).
Thirdly, and more commonly now in the UK, we can perform genotyping (but it has to be remembered that genotyping is only a prediction of what antigens may actually be expressed on the red cell surface).

The AABB Technical Manual states that antibody titers should not be performed using gel technology, so we revised our procedure to go back to tube method.
 

Hear, hear Scott.
How can an algorithm tell the difference between an anti-D+C and either an anti-C+G or anti-G, anti-hrB, rather than anti-C+e or anti-hrS, rather than anti-ce (anti-f), to name but a few?  The answer is that it cannot, and these specificities are much more common than a lot of people think.

The sensitisation stage of antibody/antigen reactions follows the Law of Mass Action.  The rate constants for the forward and reverse reactions will change both with the temperature of incubation, and also with the concentration of the antibody and antigen (amongst other things) and this will, in turn, alter the equilibrium constant of the reaction (which, in the cross-match, you want to drive to the right, but not so much that you will get "false positive" reactions).  You would be ill advised, therefore, to carry on the tests at 37oc and AHG after saline replacement, as there is every chance that the equilibrium constant would be altered to such an extent that some antibodies would not be detected in vitro, that may be clinically significant in vivo.

Sorry for the delay in response. We make up our Trypsin that we get from Sigma Aldrich. It seems to work just as well as DTT, but we treat it with both since the "standard" seems to be DTT. I've attached our Trypsin testing SOP.
Trypsin Testing.docx

All excellent books, but I have to say that Mollison's Blood Transfusion in Clinical Medicine is a real textbook, rather than a book to read from cover to cover (and I can say that with all honesty, having read it from cover to cover for a book review - and I know Dave Anstee wouldn't argue with me saying that)!

The AABB Technical Manual.

I work at a reference lab as well and we get roughly 3-4 DARA patients a month. More often than not, half are repeat customers. It seems strength varies from patient to patient, but definitely increases as the treatment continues, but nothing more than a 2+. We treat screening cells with DTT every time. We also treat a second set with trypsin to help rule out Kell group system antibodies. So far we haven't had much viability with cells lasting longer than a full shift.

We have been using pre-warming in the UK since before I started in Blood Transfusion (circa 1973) and we have never had a clinically significant transfusion reaction caused by warming away an antibody in all that time.
Yes, there have been occasions when, for example, an anti-S has disappeared by pre-warming, but, if you look in most text books, and all reliable text books, anti-S is only rarely clinically significant - and certainly none of those that we have "warmed away" have caused any transfusion reactions at all.
There was one case of an anti-Vel causing a fatal transfusion reaction, BUT, that was not missed through pre-warming; that was missed because EDTA plasma was used, and the anti-Vel could only be detected in serum (confirmed by the IBGRL), and so I think that the worries about pre-warming are vastly over estimated.
It is vital to keep up with competency for this technique, as with any other technique, but probably more so with this technique.

My pet peeve is the placement of labels - nursing will not cover the attached label on the tube so you get a specimen where you cannot see the plasma/cell interface.  Regardless of how many times we educate on this it refuses improve.

Then, presumably, he has a positive DAT due to the anti-A in his own plasma?  If so, you did not supply this information.