galvania

Hi everyone. I came across this forum by accident, while I was looking for something blood-transfusion related on the internet. I just want to say what a good idea it is. I work for DiaMed in Switzerland and before that I worked for many years (too many!!) in various labs in the NHS in the UK, where I did all my training. It's really good to be able to see different problems from different countries' perspectives - with different regulations in place and different priorities. I shall be following the threads with great interest - and maybe I can even contribute from time to time!

Dear Kyaw
What you are seeing here is surely NOT an anti-A1.  You also have a reaction with the O cells.
You should put up a complete panel at room temperature .  The antibody screen was negative at room temperature, but that does not exclude the presence of cold antibodies (for example P1, M) active at room temperature.  It may also be a mixture of a cold anti-H and another cold antibody (It can't be just an anti-H as the reaction with the A cells is stronger than that in the O cells)

First of all, please do not worry.  IF you do have anti-D antibodies, and they are real antibodies, this is only one of a number of tests that the doctors will do during your pregnancy to make sure everything is going OK with your baby.  
What they should do is recheck your blood for anti-D levels now and again in about 4 weeks' time to see if there is any change.  It will show anti-D because of the Rhogam, but the important thing is to see whether the level increases significantly over time.  Also, even now, if it is very high (VERY unlikely) then that would indicate it's real anti-D as opposed to the Rhogam.
Ultrasound is a good idea.  It is usually done anyway during pregnancy, but it will also show if something is happening that they need to react to.

yan xia, there are two mutations present.
The first, RHD 4.0/RHD DAR3.1.  This will lead to the expression of a Partial/Weak D.

The second mutation is a hybrid of the RHD and RHCE gene, with exons four to seven of the RHD gene being replaced, or substituted, by exons four to seven of the RHCE gene.

I hope this helps a little.

Also, the polyclonal (human) reagents will give false pos results in samples with pos DATS.  But anyway, finding sufficiently good human antibodies to manufacture reagents from is getting harder and harder.  So definitely monoclonal

Do keep us updated.  I am sure we are all looking forward to news of a healthy baby

Well with that sort of turnover, that sounds like a real waste of time to me as well as a horrible source of error.

This thread is pretty old but as it comes up again.... this "air gap" is required to avoid having the Anti-Human Globulin (AHG) present in the gel matrix getting partly "neutralized" by the excess of human immunoglobulin from the plasma. Keep in mind that plasma is full of various human Immunogloblins which will be recognized by the rabbit AHG. This "partial" neutralization may weaken the reaction indeed. 
Same as in tube but there is no washing step requires as the air gap is there to prevent this neutralization. Once cells are sensitized after the 37°C incubation step, the card is spun and the AHG will catch the Ig bound to the RBCs leading to positive reaction.    

How are you doing your crossmatch?  This could, as Arno said above, be an anti-buffer reaction - or it could be a cold antibody that's got enough time to stick on to the red cells before they get to 37°C.  Can't be an antibody against a low-frequency antigen - not with 4/4 being positive.  I would also double check that the blood bags really are Jka- and of the correct ABO group.  You haven't answered the question about the patient's blood group.........

How are you doing your crossmatch?  This could, as Arno said above, be an anti-buffer reaction - or it could be a cold antibody that's got enough time to stick on to the red cells before they get to 37°C.  Can't be an antibody against a low-frequency antigen - not with 4/4 being positive.  I would also double check that the blood bags really are Jka- and of the correct ABO group.  You haven't answered the question about the patient's blood group.........

How are you doing your crossmatch?  This could, as Arno said above, be an anti-buffer reaction - or it could be a cold antibody that's got enough time to stick on to the red cells before they get to 37°C.  Can't be an antibody against a low-frequency antigen - not with 4/4 being positive.  I would also double check that the blood bags really are Jka- and of the correct ABO group.  You haven't answered the question about the patient's blood group.........

Is the buffer used for preparing the RBC suspension for X-Match the same as for the AC? If not, this patient may have an additional Ab to a buffer component? 

Either the units were Jk(b+), rather than Jk(b-), or, perhaps, the patient has produced another specificity?

Sorry - I forgot the C.
As the frequency of C is about 68%, you can remove 2 of those three.  That leaves you with 1 unit
Anna

Very very roughly, and assuming your donors are mostly White European
If you start off with 100 units, approx. 50 will be O or B
Of those approx. 5 will be K+ - so you will have 45 units.
Of those, about 2/3  (so approx. 30) will be Fya+, so you will have 15 left
Of those about 80%  (about 12) will be Jka+
So, if you're VERY lucky, the remaining 3 will be compatible.
Good luck!!!!!

so very sad.  A great loss to the profession

ABO mixed field must be explained; find out patient transfusion history. If it is not clear what their blood type is, or if the mixed field cannot be explained (patient intubated, confused etc.) document and give type O.
Interpretation of mixed field in gel is easy, harder in tube but I would expect it to be there. I would therefore suggest checking very carefully for mixed field by tube (this may be an occasion to use a microscope to confirm mixed field if needed). Sounds like this is a good sample to use for mixed field training in your lab.   

It would be really useful if you could tell us the ethnicity and age of the patient, and his medication regime.
That having been said, I note that the antibody screen is positive, that his DAT is positive by both anti-IgG and anti-C3d, that the neat plasma contains an apparent anti-E and anti-c, but that the eluate contains an antibody that is, apparently, pan-reactive.
Very often in these cases, the apparent antibody specificity in the neat plasma is a mimicking specificity, rather than a true specificity.  In such cases, the apparent specificity in the neat plasma can be adsorbed out using red cells that are negative for the antigens of the apparent specificity; in this case R1R1.  The true specificity of the antibody could be an anti-Rh17 or anti-Rh18.
While I am not saying for a single second that the apparent specificities of anti-E and anti-c are not true specificities, it may be worth your while seeing if they can be adsorbed out using R1R1 red cells.  However, as you suspect the presence of other antibodies, this should not be attempted until you have proved otherwise.  This you can do, as you suggest, by alloadsorption of the neat plasma using two or three adsorption cell types.
In answer to your last question, with regard to adsorption of the eluate, this was certainly a method we used in the Reference Laboratories of the NHSBT in the UK.  It was usually used when the patient had a known pan-reactive autoantibody, but was requiring transfusions more frequently than previously, and/or when the expected rise in the haemoglobin concentration was not achieved.  On some occasions, we were able to detect a de novo alloantibody in the eluate that we could not detect in either the neat plasma, or the adsorbed plasma, although this was not always the case, as transfusion in and of itself can sometimes stimulate the autoantibody to become more active (see Petz LD, Garratty G.  Immune Hemolytic Anemias.  2nd edition, 2004, Churchill-Livingstone).
Good luck with sorting it out, but this is a really interesting case.  Thank you for posting it and, please, would you mind letting us know how you get on?

..........and on the result of the reagent control you put up with it and what is wrong with the patient, if it's a man or a woman,and how old................and why you were doing the test in the first place

Or was the K_B positive because mum had high levels of HbF and therefore none of the injected anti-D 'used up'?......
Butlermom - where are you??????

..........and on the result of the reagent control you put up with it and what is wrong with the patient, if it's a man or a woman,and how old................and why you were doing the test in the first place

I know that some of the early work on ABO-mismatched solid organ transplantation, viz-a-viz ABO antibodies was carried out by Professor Patrick Mollison and his co-workers, and he showed that, whereas inhibition of IgM ABO antibodies is reasonably easy by, in the early days, transfusion of FFP to adsorb the antibodies in vivo, the same is not true of IgG ABO antibodies.  He and his co-workers found the inhibition of these antibodies was much more difficult, and this was almost certainly because only 40% of IgG antibodies are intravascular, as so they "rebound" when inhibited or removed from the intravascular area, whereas almost all of the IgM antibodies are intravascular, and so "rebound" is less likely.

Not only is it possible, but your supervisor has done exactly the right thing.  They other techniques are a complete waste of time and money that tell you precisely nothing of any worth.

oh my goodness.  You poor thing.  You have really been through it.  I admire you for your strength.  You are very brave

I would just be a bit suspicious if say there was a + reaction with A1 cells and 4+ with B cells in an apparent group O - or vice versa; or very weak reactions in a young healthy adult.....a bit of common sense required, that's all