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Bb_in_the_rain

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Everything posted by Bb_in_the_rain

  1. Yan xia- I have no idea why it does not react with a 6 month old frozen/thawed AB plasma but reacted with fresh AB plasma. I treated 3 more sources of cells and they reacted weaker (2+) with this same source of frozen/thawed AB plasma but strong (4+ blasted) with fresh AB plasma. May be anti-T diminishes with storage? Galvania- I think docs are suspecting an infection in this baby.
  2. Hello. sorry about the delayed reply. We worked further on this case and turned out the pooled AB plasma that we have frozen did not react with neuraminidase-treated cells (another aliquot of NeuNac treated cells) but fresh AB plasma did. We did test this patient's cells with fresh AB plasma and got a positive reaction. We have to try not to used frozen/thawed AB pooled plasma for this test in the future
  3. I read in Dr.Garratty's Immune Hemolytic Anemias textbook that Spontaneous agglutination is often seen with warm IgM cases. Would it be helpful to spin this patient's cell suspension with either saline or 6-10% albumin if the warm IgM AIHA is in question? And also preform titer and thermal amplitude test to see if antibody titer increases or decreases with higher temperature?
  4. Are your cells that is non reactive with eluate either e- or f-? did you have antigen typing on this patient? Was the patient recently transfused?
  5. I recently used this procedure to differentiate anti-Pr from anti-En(a), which is the only time I have used it. Since our lab refer possible Asub for genotyping lab, we have no chance of trying this out to catch weak subgroup of A.
  6. Hello, I am wondering what kind of method do you use to quantify anti-D in europe. here, we titer the maternal plasma with D+ cells and freeze an aliquot. Next visit, we would titer the fresh plasma parallel to thawed plasma, see if the titer has gone up compared to previous titer. Seems like having an assay to quantify the amount of anti-D and having a clinical cut off correlation is a better way to approach this.
  7. Anti-CD38 did not react with In(Lu) cells and showed very low signals when anti-CD38 was tested with In(Lu) cells in a flow cytometry. Before we knew the patient was on anti-CD38 and In(Lu) cells has depressed CD38 marker, we have mistaken it being an antibody to high incidence antigen in Lutheran blood group system, based on the reactivity with In(Lu) cells.
  8. Looking at this kid's clinical information may be a good start. What is the diagnostics? is the kid actively hemolyzed? What is the medication history? did you try washing this patient's cells with warm saline and try ABO/D typing with warm-washed cells? That is feasible to do in hospital lab. I would even try using DTT if you have some in-house.
  9. Hello, I am learning so much from you guys here as usual. I also realized this english flag along with the 2+ IgM on the DAT result. I started wondering what kind of direct antiglobulin test is performed to detect anti-IgM in the UK. Do you have liquid antisera (monoclonal, animal antisera etc) or anti-IgM suspended in gel?
  10. Beautiful pictures. Thank you for sharing the cases. I have been on this site for almost a decade and never stopped learning from you guys! I wish we have these gel cards here in the States. There were sometimes we suspected problem due to IgA in DAT negative patients and those cards will be great to investigate such cases. I tend to attempt to "reinvent the wheel" due to my lack of historical knowledge and experience. I wonder if the infants cells from moms with Kidd blood group antibodies screened with anti-C3 in the past to see how many of them have turned up positive. It will be an interesting data to see if that has not already been done.
  11. I am wondering why the cord cells were tested with anti-C3? I believe it is not a common practice to test the cord with anti-C3 I just saw one too a few months ago, mine is a peripheral draw on a new born rather than cord, anti-IgG and -C3 pos. I looked it up on Issitt's textbook. His references indicates that HDN due to complement activaton is an argument that has been challenged, as "C3 coating can be found on normail cord samples". I think if Hb and bili are normal C3 coating probably is not significant??
  12. Very interesting case! Here are my questions related to this case. 1) If it is a case of auto-anti-LW, should we expect the autoantibody to have a "relative specificity" to D antigen, especially with a 3+ reaction by ortho gel. Would a transient suppression of LW antigen effect D antigen typing by a monoclonal reagent? 2) Antigen loss during AIHA is shown in Kell blood group system. Is it possible in Rh blood group system as well? Zimring JC, et al. Antigen loss from antibody-coated red blood cells. Transfus Med Rev. 2009 Jul;23(3):189-204. PMID 19539874
  13. here is a fun case along with a question that I cannot wrap my head around it. Mother: Anti-D and anti-G identified (both strongly reactive by PeG and saline-IAT), Anti-C ruled out by differential adsorption and elution with R2R2 cells and r'r cells. All other common alloantibodies ruled out. She was not previously transfused or pregnant. TITER with R2R2 cells = 1024, titer with r'r cells - 128 (I know this titer is so unhelpful since expression on each of our indicator cells are different) Baby: (here is the fun part.. ready?) - on the birthday D typing negative with MoAb anti-D reagent at room temperature with untreated cells, EGA treated cells, twice EGA treated cells. DAT 4+ with untreated cells, EGA treated cells and twice EGA treated cells. Baby is fine (dont need transfusion, not hemolysing). Plasma and eluate reactive with D+ and C+ cells. insufficient sample to perform adsorption/Elution to differential anti-D from anti-G. eluate- reactive with D+ cells and C+ cells/ we assume it was anti-D and anti-G in there. Genotyping result. RHD*DAU0. Homozygote D deletion detected. So baby is heterogygote DAU-0 without a normal D gene. 1 month later, baby came back hemolysing. DAT 4+ with untreated cells. Anti-D typing 4+ with EGA treated and untreated cells. DAT on EGA treated cells is negative. plasma and eluate reactive with C+ and D+ cells. all other alloabs ruled out. Not enough sample to perform adsorption/Elution My question is.. The baby's red cells were clearly coated with anti-G and/or anti-D why did it not hemolyse at birth but 1 month after. I have heard of blocking mechanism serologically as in this case Does the heavy coating of antibodies makes Fc receptor inaccessible for macrophages for hemolysis? Could it be anti-G alone (not anti-D) coating on the red cells protecting the red cells from hemolysis by anti-D?
  14. Ours is DTT. We tried cords (alot of different sources) Not every cord cells gave is a negative reaction with plasma from patients on DARA. half of them still reacted microscopically.
  15. Thank you very much! This is very helpful to see that cells that are previously frozen in liquid nitrogen can be transferred to -80C and it would recover well. I should have been more clear in my question (#2) above. What I envisioned was making droplets in liquid nitrogen and storing the vials with droplets (instead of an aliquot) to avoid having to thaw and refreeze the same vial every time we use the cells. The plan was to take one frozen droplet out of the vial and use it instead of thawing the entire vial to take out a drop. I assume Alan's statement about previously frozen in liquid nitrogen and recovering well in -80C storage implied that we can make droplet in liquid nitrogen using glycigel and store them in -80C long term?
  16. I am reading the same book right now and am very happy to learn about a history of how this field is eevolved and the key players in the field. Also, realized I was born a little too late. It would have been nice to live in the era of exciting discoveries.
  17. When we have these in our central transfusion services, we look into it a bit deeper here in IRL. We repeat tube method using 3 different sources of anti-D. If all 3 sources were negative, report RhD negative. If all 3 sources were >2+, report RhD positive. If 1 of 3 sources were <2+, refer to genomic testing looking for WeakD type 1, type 2 or type 3 (if patient is female childbearing age or <18 year old). Genomic WeakD type 1, 2 or 3 were treat it as RhD Positive. If not genomic WeakD type 1, 2 or 3, treat it as RhD negative. If 1 of 3 sources were <2+, all 3 sources give positive reactions and patient does not have anti-D and patient is not female of childbearing age or pediatric (<18 year old), report as RhD positive. (if the patient, later on, makes anti-D change it to RhD negative) Phew.!! I think the decision overall is more based on the patient's age and demographic.
  18. Thank you very much. Cant wait to play with this glycigel
  19. Hello, Unfortunately, I could not make it to AABB meeting last year. Any update on this? I can say that our lab alone here would want 10+ copies here. Please let me know if you would like to make up an online survey or poll sheet that a link can be shared around. Also, would that be helpful if I ask sale specialists from commercial blood bank supplies on site here about sponsoring for the printing. Would you mind giving me some details about how to go about it? "Books created after Jan. 1, 1978, regardless of whether they’ve been published, are protected by copyright for the life of the author plus 70 years. Some books that are works-for-hire, i.e., created in the scope of employment, or that are created anonymously or pseudonymously are protected for 95 years from the date the work was created or 120 years from the date it was published, whichever is shorter. The law on copyright duration has been amended many times and determining whether an older book is still protected by copyright is more difficult. However, any book published prior to 1923 is currently in the public domain. Works published prior to Mar. 1, 1989, without any notice of copyright are also in the public domain, because the law required notice on all works published before that date." Does it mean reprinting this book is out of question?
  20. I would like me a third ed if it were to go in recycle. Is it still available?
  21. There is someone selling digital copy for $500 each. omg! https://www.amazon.com/Applied-Blood-Group-Serology-Issitt/dp/0935643052/ref=la_B001KCKXFY_1_1?s=books&ie=UTF8&qid=1506652837&sr=1-1
  22. Just worked on some. Please email me to dothandar@gmail.com for more information
  23. Opps my appology. I just saw the recipe for "Brentwood" solution for thawing.
  24. Plasma from patients from DARA is non-reaction with In(Lu) cells I think. CD38 maybe suppressed on In(Lu) Cells? May be Kell blood group antigens excluded using In(Lu) cells that are K+k+? Or cord blood?
  25. Thank you very much for the explaination. It is very helpful. We are very lucky to have a resourceful mentor like yourself who is here to answer any questions that we have. I am checking out the recipe of glycigel in the library and I have a couple of questions. 1) What ratio of red cells to glycigel is appropiate? Do you warm up the gel before suspending red cells into it? If so, we have to keep the red cell suspended gelatin warm till we freeze the sample? 2) Can I make red cell droplets in liquid nitrogen using glycigel suspended red cells and keep the vials in -80C freezer? (since we do not have long term nitrogen storage here). 3) How would I thaw the glycigel suspended frozen red cells? Do I just suspend it in warm 0.9%saline?
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