missyg

Because the technical Manual does state packed cells we now use donor cells for DTT treatment. We have found that we get better results than the DTT treated reagent cells. The reagent cells seem to lyse more and we have had to repeat testing a lot because of that.

We performed a prewarm Lo Ion panel and Peg panel.The results were: 1 out of 3 M homozygous cells reacting and 3 heterozygous negative. Yes we are testing in tube. The titer for monitoring maternal specimen was performed using no enhancement and was negative at 1:1. At this point the anti-M appears to be IgM but the DTT does not seem to concur.

Pregnant patient with an Anti-M reacting through all phases(immediate spin, RT, 37oC and IgG). 4+ IS, 4+ RT, 3+ 37oC, 2+IgG with homozygous M (heterozygous is slightly weaker). 37oC and IgG was with Lo Ion enhancement. .01 M DTT treated serum and PBS control reacted same strength indicating incomplete denaturation of IgM class. We had some suspicion that the DTT was not working however we had used the same DTT on other patients and it worked. Could it be the strong reactivity of the Anti-M? We performed some titration studies using neat plasma/serum, the DTT treated serum and PBS control with results as follows: Neat: IS 32 30 min RT- 128 37 Lo Ion-32 and IgG- 16 60 minute 37 incubation IgG phase no enhancement: <1 DTT treated no enhancement IgG: <1 PBS control no enhancement IgG: 32 Any thoughts are appreciated!!!!!!

We have recently had a donor who has donated multiple times and come up positive on automated (solid Phase) antibody screen, positive in our tube antibody screen and negative in the tube antibody panel using murine monoclonal IgG. The last time the donor came in we ran the antibody panel in gel and there was a panagglutinin with autocontrol positive and Polyspecific gel DAT positive. We performed a 2x 37C allogeneic adsorbtion and ran the adsorbed plasma in gel. The adsorbed plasma again reacted same strength no change. We then made a connection that the IgG card for Ortho is Rabbit IgG, the automated IgG is Rabbit and the ABS tube method IgG is rabbit. We performed the tube antibody panel using Rabbit IgG and it too is positive now! All the same cells that were negative with the murine monoclonal IgG are now positive with the rabbit IgG. Somewhere I have read about the Rabbit IgG containing IgG4. Could this be a IgG4 warm autoantobody? Thanks in advance!!!

Well we knew that something was happing to "neutralize" the IgG portion. We performed a mini cold and found a cold agglutinin present. We then decided to wash the cells with warm saline and it worked! We believe there was some kind of protein present that was neutralizing the IgG that was only removed by warm saline. But we have our result! Thank you to all who responded!

Yes we perform QC every day! It was also washed multiple times by different methods and it still does not check cell. Again it is only the IgG portion that is not working because we used the complement check cells with the polyspecific and it worked. Also to note that the DAT in gel was 3+ with Polyspecific and IgG cards but appears negative in tube (but not valid due to check cell failure)

DAT workup on a patient. At Hospital weak DAT positive (microscopic positive) IgG and Polyspecific post transfusion. We have performed the DAT in tube method and the complement and saline control were negative and check cell worked but the IgG and Polyspecific were negative (note we do Not read microscopiclly per Technical manual) but the IgG check cells failed on multiple attempts. We hand washed and used the automated cell washer and all check cells failed. Just out of curiosity, we used the complement check cells for the polyspecific and it worked. We do not have too much information on the patient. We have a few hypothesized reasons as to why the IgG seems to be neutralized but nothing concrete. Any suggestions?

Thank you all for the suggestions!!! We are trying some other techniques and contemplating having the patient genotyped. As far as we are told the patient was not transfused anywhere else. The patient does not have bacterial infections and is awaiting liver transplant. I will update you all as soon as we have results!

Patient is 58 years old

Has anyone observed anti-Jka in an eluate of an individual who is Jka negative and received only Jka negative units? If so, can you offer an explaination? Thanks!

Thanks!!!!!

We have a patient who has the known antibodies: anti-Fya,-C,-K,-Jkb. The patient now has a cold agglutinin and a warm auto antibody. The patient's race is black and phenotypes as Fya and Fyb negative. A 2 X @37C allogeneic adsorption was performed using R1R1 Fya+(cell A), R2R2 Fya-(cell , and rr Fya+(cell C). Given the patient's race, we wouldn't expect them to have an anti-Fyb. However, when the adsorbed sera from cell A and C was tested with Fyb positive cells it reacted 1+. If this were anti-Fy3, we should have expected it to be adsorbed out with the cell A and C and in turn have no reactivity with the Fyb positive cell (as a side note, the Fyb positive cell was negative for all the antigens to the previous known antibodies). All other common alloantibodies have been ruled out. Any ideas what could be causing the reactivity with the Fyb positive cells??????????????? An autoadsorption was also performed supporting the results of the allogeneic adsorption. Thanks!!

We have a donor who was a previous patient and has Anti-Yta,-Fya. We have to redemonstrate the previous antibody and rule out common alloantibodies every time the donor comes in. My question is, if we rule out the common alloantibodies by adsorption (redemonstrating the Fya) and the neat panel is positive (negative auto control) demonstrating the Yta, do we also have to run the Yta and Fya negative cell everytime to prove it is indead the Yta and not some other high incidence antibody? I just worry that we may run out of the rare cell as this donor is in frequently. Just curious for other opinions! Thanks!!!!!!!!!

GW is George Washington and it is a review course that is 12 weeks long. And I am missyg!!! There are also some really good study aides in the library section of the blood bank talk forum.

George Washington University has a good SBB examination review course online!