trisram
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Posts posted by trisram
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Forgive me if I am being a bit stupid here. Antibody screening and identifcation is carried out by adding patient sera to unsensitised screening cells and panel cells in BLISS/LISS, incubating and then performing Indirect Coombs tests (IAT) on each tube. The DAT (direct antiglobulin test) is used to detect antibody allready attached to red cells usually the patient. In other words the IAT technique is the first test used in detecting antibodies. Correct me if I am wrong.
Regards
Steve
:confused:You're correct, IAT means indirect coombs test, not indirect antiglobulin test ... Sorry, I forgot what IAT meant. I see what you mean.
Sorry, I usually work in microbiology. I am no blood bank expert.
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Apologies accepted, I hope you mean you use 'Indirect coombs' (IAT) not direct coombs
Steve
:)No. Direct coombs, DAT, in vivo. I don't know anything about IAT .
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'nevermind. thanks. what kind of panels you use? 11 cell panel, all O Positive cells? or do you use select cells too? any other reagents? thanks, but this doesn't answer my question one bit['
I am also sorry that I have not been of any help. However, I will answer some of your question. I use a 10 cell ID panel which when you include the 3 screening cells both suuplied by the UK Blood Transfusion Service makes 13 cells in all. Screening is carried out using an IAT technique, the Antibody ID panel includes papainised and a IAT techniques.
What Malcolm described was more thorough than ny input and is based on the minimum requirements for antibody identification as described in the UK Guidelines which you will find in the reference tab of this website.
I have to say trisram, I am now at a loss as to what you want to know and the attitude you have shown in your reponse does not encourage anybody to offer any help to you.
Regards
Steve
:confused:I apologize. We don't use IAT here where I work, just the direct coombs. But anyways, I was out of line. I apologize
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Obviously, my answer did not help you; even a little bit.
I must apologise to you for writing rubbish.
I will leave responses to any other of your posts on any thread to other people with a better understanding of both what you are asking and basic serology.
:mad::mad::mad::mad::mad:
I apologize, I was out of line.
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Dear trisram,
We are only a small laboratory, but even we can resolve a lot. The antibody ID panel we have in use will resolve most of the significant mixtures of two antibodies. Once we have reached a conclusion then we test the patient red cells for the corresponding antigens to confirm that the patient could be stimulated to produce the antibodies.
Then finally. we send fresh samples to the local RCI laboratory (i.e. Malcolm's lab) for confirmation
Regards
Steve
:)nevermind. thanks. what kind of panels you use? 11 cell panel, all O Positive cells? or do you use select cells too? any other reagents? thanks, but this doesn't answer my question one bit
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The answer to 3 is that you would use 2 (or more) cell samples positive for the antigen against which you think one of the antibodies is directed, 2 (or more) cell samples positive for the antigen against which you think the second of the antibodies is directed, and so on, and 2 (or more) cell samples that are negative for all of the antigens against which you think all of the antibodies are directed against.
In addition, as my very good friend Mr. Jeff so correctly says, you should type the red cells of the patient for the antigens against which you think he or she has made antibodies.
4. As long as your assumptions are correct, order your units and cross-match them!!!!!
As for the methodology, you must use, as a bare minimum, the method by which you first discovered the reactions (i.e. however you did the screening and initial antibody panel) and, if by any chance this did not include an IAT (although I cannot imagine that it would not), an IAT.
Hope that helps, at least a little.
:):)your answer doesn't make any sense. where are you getting all these cell samples? so I get both positive and negative cells? what does that prove? can i just expired panel cells as select cells?
I just want to rule out everthing but my two suspected antibodies. that's it. I don't need all these other rubbish
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The answer to 3 is that you would use 2 (or more) cell samples positive for the antigen against which you think one of the antibodies is directed, 2 (or more) cell samples positive for the antigen against which you think the second of the antibodies is directed, and so on, and 2 (or more) cell samples that are negative for all of the antigens against which you think all of the antibodies are directed against.
In addition, as my very good friend Mr. Jeff so correctly says, you should type the red cells of the patient for the antigens against which you think he or she has made antibodies.
4. As long as your assumptions are correct, order your units and cross-match them!!!!!
As for the methodology, you must use, as a bare minimum, the method by which you first discovered the reactions (i.e. however you did the screening and initial antibody panel) and, if by any chance this did not include an IAT (although I cannot imagine that it would not), an IAT.
Hope that helps, at least a little.
:):)Thank you, but what do you mean, 2 from cells samples? where can I get these samples? from the cell sample store? or can I use prior expired panel cells that are positive/negative for the corresponding antigens?
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3 is not a question
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1) You get a postive antibody screen.
2) You then do an antibody ID panel.
3) The results of the panel makes you suspect 2 or more antibodies in the patient's plasma.
What do you do next and what reagents/methodology do you use?
Thank you in advance for your time and help!
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Hi trisram,
If you don't have an appropriate answer to my question, I guess you have the right to remain silent.
I hope nobody has given you the authority to make decisions on me.
Moreover,my advice to you is; please mind your own business.
And if you want to reply to any member's question in the future, please reply to the subject. Don't try to make decisions on them.
I wasn't trying to make decisions on you. I apologize if it seemed that way. I am just tired of incompetent people working in the medical industry. Enough patients have died already.
Nothing against you personally because I don't know anything about you. But, anyways, I apologize, so sorry.
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there are 2 diluents in the US ... MTS 2 and MTS 2+. The 2+ is for cells not collected in EDTA.
I don't care about diluents. This still doesn't answer my initial question. Dude, you're a SBB, do you know the answer?
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At my facility I created an SOP called Anitbody Indentification Guide ever since we went to gel and had all these problems. What we do if we see Mixed Field in gel is repeat the antibody screen using 3.0% cells and continue on using tube method. i.e Saline replacement is used for determining Rouleaux, it is not a method for crossmatching. Or Cold agglutinins. BE CAREFUL about using WARM Saline for saline replacement, you could be washing away an antibody. ROULEAUX is due to the high proteins in the myeloma patient and is a nuisance but must be careful of your methods as not to miss antibodies. Patients who react in gel due to your discribed problem we tag them to be done by tube method. This has cut down on repeat problems each time they come back for transfusions.
Thanks but you misunderstood what I said. The washing with warm saline procedure was from the people who makes the gel cards. It wasn't for replacement, because there was no plasma to replace and no antibody to wash away. We add the plasma afterward.
Well, this is what happened:
1) We did the gel card Ab screen and it showed rouleaux in the gel.
2) we read manufacturer's procedure of ridding rouleaux by washing screening cells 6 times
3) after washing screening cells, we pipet them into the gel cards and finally add the patients plasma
4) incubate for 15 mins
5) centrifuge gel card
6) Rouleaux all gone
My question is , why wash the screen cells? what are we washing off?
forget the plasma. this is not replacement. we add the plasma after we wash our screen cells with warm saline? why warm saline? why not cold?
like I said, if Malcolm doesn't know the answer, chances are, nobody does
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Wow, if Malcolm doesn't know the answer, it probably doesn't exist
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I would wash and resuspend with diluent 2. There is another thread about concentrating .8% cells and I posted I had switched from a .8% panel to the 3% panel. . One of the reasons I switched was we had a couple of cases where we were getting reactions with all our .8% screen and .8% panel cells and referring them to our reference lab. They didn't get anything. Since our autos suspended in diluent 2 were negative, I decided to try diluting a 3% panel with diluent 2. Whatever the reason, we have gotten a few patients where the screen cells are positive, but the diluted 3% panel is negative. I first noticed this problem after the new formulation for the .8% cells so I totally agree with Malcolm that it might be the antibiotic they use now in the .8% cells. On the ones where the diluted 3% panel was negative, I did try washing the screen cells with the diluent 2 and testing them and it made the reactions weaker. Since I had done a gel and PEG panel and there was no reactivity in either of those, we just called it a reagent problem with the .8% diluent.
which one is diluent 2? is that the mts diluent?
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Is it acceptable to wash the cells with saline and then use the cells on gel? The insert for the 0.8% says to use the cells directly from the vials. If you are washing with saline, aren't you washing off the enhancement?
what enhancement? I thought that was the clear liquid in the gel cards
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I had this patient with multiple myeloma. Her antibody screen on gel card had rouleaux reactions for all 3 screening cells. I then washed the screening cells six times with warm saline. The screen was then negative.
Forgive me if this is dumb, but why wash the screen cells? Obviously it worked, but isn't rouleaux a plasma property? That is why we do saline replacement when doing tube cross matches.
I was just wondering if there is a mechanism or principle I am missing here. Thank you for time and reading.
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I am looking for information on frequency of antigens. Like, which are most common and which are not. Can anybody help me find this information? Thank you for your time
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I was just wondering why is important to test for little c antigen on all donor units before issuing the units to a patient who has Anti-E.
Does anybody else do this? Thank you for your time
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Each time FFP is needed, if a type&screen is not done within the last 3 days, FFP won't be issued. Once the type&screen is performed and the screen is negative, that is when we issue the FFP. If the screen is positive, we won't issue until the Ab ID is done.
I've never been in a situation where there was no time to do a type&screen just to issue FFP. Usually, in emergency situations, blood is issued.
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Well, there won't be a need to do a gel crossmatch if the patient's screen is negative. I was talking about the screen mostly, since rouleaux is mostly a property of the plasma, not cells. Of course washing cells is mandatory and important. Anyways, everyone always wash the patient and donor cells.
If having to do a gel crossmatch, take the extra couple minutes to WASH the donor's cells freeing them of the proteins and fibrinogren that may cause rouleaux but mostly freeing them of storage byproducts.. Experience has taught me that spending a little extra time in preparation will eliminate the need to troubleshoot and worry later. -
You don't need to do Step #4. If the DAT is Positive, you need to figure out WHY. Is it due to an ABO antibody or due to some other (ie: "unexpected") antibody from the mother? (Sorry....I forget what ABO/Rh you said the baby and mother were.) (But there is no reason to prewarm, wash, or repeat the DAT.)
You probably assumed I washed the cells before I tested them with AHG. The DAT could be a false positive. You need to wash cells because there may be Wharton jelly on it.
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A 60 year old man is admitted into the ER. He has a history of colon cancer. Some blood is drawn from him for lab tests:
His blood culture came up positive for E. coli.
His hemoglobin is : 8.1 g/dl
His ABO RH is:
Anti-A= 4+
Anti-B=0
Anti-D=4+
a1 cells= 2+
b cells= 4+
What could be the explanation for this discrepancy?
Here is what I answered:
Patient possibly have sub-group other than A1 cells, perhaps A2.
I test commercial A2 cells with patient's serum. Then test patient cells with anti-A1 lectin. If both tests are negative, then patients cells are A subgroup other than A1.
So his blood type is A+.
Did I miss anything? I am just wondering why the E.coli part was added to the question. It is known that gram negative bacteria can modify the A antigen to a B antigen, and I was thinking maybe this was a B antigen phenomena question, but then, that would have mean, forward typing would have been positive for B antigen. Maybe that E.coli thing was thrown in there to confuse things. Thanks.
:confused:
:)
MT
in Transfusion Services
Posted
can you send me too? rsnowayinhell1@gmail.com