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David Saikin
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Posts posted by David Saikin
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Mabel
I validated all my rares using gel cards, either buffered or IgG. I use 50uL of 0.8% cells and 25uL of the antibody. These are incubated as per the pkg insert (room temp, 37C) and spun for 10 minutes in the MTS ctfg. The only rare that didn't validate was my 4 yr outdated anti-Lea. This includes the monoclonal anti-K1. Did you say monoclonal Jka? If yes, who do you get it from? By-the-by, the Immucor complement check cells react 3-4+ in gel (after 5 min incubation). An even more efficient way to use antisera is with Chown tubes. You can do about 50 types with 1 drop of antisera. We used to use them to screen donors for "e". Of course we had to verify using the manufacturer's directions, but a 3 mL bottle went a long way.
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An antibody screen is not required by Standards post-partum.
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Irradiate to prevent GVHD.
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I buy small manilla tie tags. The tech tests the units when indicated; the tie tag is stamped with our blood bank identification on one side. One of the unit bar code labels is placed on the other side along with the antigen typing, date and tech. This is then tied to the bag. The unit testing and control results are recorded in a manual paper log (but will soon be a part of a computerized testing record).
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We use anti-IgG gel cards and buffered gel with anti-C3b,-C3d. We use both for adults and cords. The Immucor complement check cells are always 3+s - 4+ in the gel card. I also run the patient cells neat in the buffered gel (0.8%) to make certain I am not seeing any non-specific agglutination (yes, that is my paranoia easer).
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If you are ging to go with an automated system - Provue, Echo, Galileo, Tango - see them all in operation and talk to the users not the vendors. If you want to do manual -gel vs solid phase, - I think you'll find the gel is more flexible, ie, you can do more things easier. I went to gel for a number of reasons - most notably it was easier for my personnel - to learn and to read. I had demos of both. An additional plus was that there were no solid phase users nearby in case I needed to "borrow" something. Everyone I know who has an automated system really likes the one(s) they have. I also have been to a considerable number of transfusion services for professional reasons - I have only seen solid phase in one.
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The ortho prices are going up 100% for all non-gel reagents. This will be effective in March sometime . . . I got this word last week. Both companies appear to be tyring to force the issue on both automation and their "new" technologies. Their justification, as I understand it, is that they are losing money with the classic blood bank reagent business. Both tried to divest themselves of such but the FDA would not allow it. About 4 years ago classic prices started to increase 100-300%/year in the corporate attempt to remain profitable. My superiors were rather perturbed at this announcement. We have no alternative but to increase our prices for antibody id, ag types and other reference work. Ain't America Great!?!?!
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We always sent a letter to the donor's physician (SOP/autologous only). It was incumbent on the MD to report any reportable condition.
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If I recall my complement activity, doesn't C3b deteriorate into C3d in vivo? It seems that a reagent that detects both will provide more information than the alternate.
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I qc my panels upon receipt. 0.8% cells are checked with a 1:10 dilution and 3% cell are checked with a 1:4 dilution (I use anti-c). Cells are visually examined each day-of-use.
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Theoretically, anti-A,-B,-D have to be checked each day of use. Tha anti-A and anti-B only need to be qc'd with a + cell. The anti-D must be checked with a + and neg cell. Also, if your lab does sufficient numbers of ABOs, the front and back types may be used to verify quality performance on that day (you don't have to perform actual QC). You determine what is "sufficient". I have a problem with QC of one bottle of a lot/day. If you are a large BB, you may have multiple racks in use. I believe that each rack should be quality assured, not just the lots. (personal bias).
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We always switch back ASAP after the emergent situation
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We use the anti-C3b,-C3d . . . why? because we always have (I guess).
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I concur with your interpretation (and I do AABB and CAP assessments). I have never seen a contract with a reference lab. We use the red cross for our blood supplier and will utilizie their reference service if something is beyond our capability. I do work for other institutions; they just need our CMS certification for documentation for their inspections.
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Yes Mabel, I run the check cells in a parallel microtube at the same time. I also run both the patient and the check cells with Dil 2+ as a negative control. I just feel better that way; due to the increased sensitivity of the gel, I wouldn't want some non-specific agglutination giving me a false +.
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Shily
Weak D+ individuals CAN produce anti-D. It depends on the nature of their D antigen. If they are missing a piece of the D mosaic, they can product ab to those portions they are missing. I would expect that Del individuals that fit that description would also be D antibody makers.
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Bob
I am with everyone else . . . thanks for all your insight and best of luck with your renewed endeavor in transportation.
Dave/Littleton, NH
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We are due to be inspected by 10/17. Last Friday, CAP called to tell us that we will be done between 10/18 and 11/17. They have a letter stating that your accreditation does not expire. The hospital 20 mi north of us was inspected on the last possible day. The hospital 20 miles west of us was done on the 2nd day of the 6 month cycle. you figure it! CAP also said we would get a call about one hour before the team arrives.
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I validated anti-C3b,-C3d using buffered gel cards. We use 0.8% patient cells and we make the commercial Complement Check cells 0.8% too. We spin after a 5 minute incubation at room temperature. The check cells are always 3-4+. As for the sensitivity problems being discussed - negative tube vs + gel . . . why are you using gel? Up front we all know it is more sensitive. I don't think you can ignore a + gel result just because you repeat it in tubes and it is negative. My ped docs are enthralled with the increased DAT sensitivity. I have found 3 anti-Jka in gel that were negative with tube/PeG testing. Are they clinically significant? I can't ignore them.
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I was fortunate enought to take the exam when it was both written and practical (I challenged the test/not SBB school). Then, the written was BB trivial pursuit. Know ag frequencies so you can calculate the % of compatible units based on these (and don't forget ABO compatible too, sometimes the questions are worded tricky and will include ABO, but sometimes they won't). On my test, we were the first to have patient management questions, as they were phasing out the practical - I found them obscure and very subjective. Donor stuff had to do with both testing/understanding cutoff # and how it was generated, donor reactions and donor suitability. Do you need to observe donors being drawn? I don't think so. Needed to know how to calculate cryo dosage/ffp factor dosage. Also understand red cell dynamics. The test was not easy. I used Mollison, Pittiglia (Harmening), the Technical Manual, and copious flash cards. As one of my techs asked me once "should she take the test". I told her go for it. As long as you can afford to lose the money, even if you do not pass, think of all the BB knowledge you will gain and be able to use. GO FOR IT!
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You might want to try setting up a monocyte monolayer assay . . . you can see if the patient's monocytes will "attack" red cells sensitized with the antibody. Someone else will have to help you with the particulars, I have only read about this assay.
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TP is not a licensed entity, unless you are using the blood for transfusion purposes. I would say the feds may still want to see records for it - they like to see that the people doing it are trained and not drawing too much. I do not have it on my registry - the feds look at my paperwork and check my qc stuff. As long as your Medical Director is responsible it should be in compliance with AABB and CAP.
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I validated my buffered cards with 50uL cells/25 uL antisera.
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I wonder what company's gel you are using? In the USA there is only one vendor. I know that in Europe there are at least 3 different suppliers. I have validated typings in gel using either the buffered gel cards or the anti-IgG cards available here. The only difficulty I encountered was with Lea typing (probably had to do with the antisera being outdated for a few years more than the gel). I did not need to use enzymes. Are these recommendations based on your personal validations or from your gel manufacturer(s)? Thanks for your valuable input.
DATs
in Transfusion Services
Posted
Mabel - I do not run an auto ct with my antibody screens. We only run one now if we are doing an identification. I am not doing any more elution studies than I have in the past, but then again, I am not performing an auto. It has been apparent with gel (from the time I've started using it) that it is significantly more sensitive than tubes - any reactivity of 2+ or weaker in gel is usually negative in tubes. Why anyone runs a more sensitive test and then verifies it with one significantly less sensitive does not make sense (to me). Why use the more sensitive method to begin with (rhetorical)? It seems that if the method you use demonstrates a detectable level of reactivity, it would be incumbent on you to follow up that detectioni. Anyway . . . my peds docs like the extra sensitivity found in gel - they thought we were missing too many +DATs.