What do you do if you suspect 2 or more antibodies?

[trisram]

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!

[Steven Jeff]

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

:)

[Malcolm Needs ☆]

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!

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.

:):)

[trisram]

3 is not a question

[trisram]

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?

[trisram]

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

[trisram]

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

[Malcolm Needs ☆]

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:

Edited April 4, 2010 by Malcolm Needs

[Deny Morlino]

Malcom,

Your explaination was helpful. I have a student and this brief synopsis helped her add to her understanding of a logical approach to identifying more than one antibody in a sample. Sometimes we get so deep into an explaination we can lose track of the goal. This helped her focus on the primary points.

Thanks,

[tbostock]

trisram, no need to be rude here. We all do our best to help each other out. Malcolm gave a perfectly understandable and helpful answer, if you need clarification on your original answer, please just ask.

[Steven Jeff]

'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:

Edited April 4, 2010 by Steven Jeff

[trisram]

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.

[trisram]

'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

[Steven Jeff]

Apologies accepted, I hope you mean you use 'Indirect coombs' (IAT) not direct coombs

Steve

:)

[Malcolm Needs ☆]

I apologize, I was out of line.

Apology gratefully accepted.

:):)

[trisram]

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 .

Edited April 5, 2010 by trisram
I took out all the vulgar words and obscenities

[Steven Jeff]

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:

[Deny Morlino]

That is the order in which we proceed. I guess I am a bit confused.

[Malcolm Needs ☆]

Me too.

:confused:

[trisram]

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.

Edited April 5, 2010 by trisram
I removed all the curses and profanities

[Deny Morlino]

Sorry for the confused reply, but I was having a hard time figuring how to perform routine blood bank workups without the IAT method. Please don't think I am picking at you; I just wanted to be sure any replies I posted would be relevant to your situation. Does your facility have a tech tasked with Blood Bank as their primary responsibility? If so, they can probably shed some light on the process most appropriate for your situation. If you are the lucky soul now saddled with the responsibility, this group is an excellent resource. Let us know how we can help!

[rravkin@aol.com]

Trisram,

I think that what Malcolm and others were trying to explain is that after the first panel is complete and you can not rule out two antibodies you must then continue with selected panel cells of panels in your blood bank other then the panel just used. These selected panel cells are cells that you select that are negative for the corresponding antigen of one of your two suspected antibodies and positive for the other; and vise versa. Here's an example, your ABSC is positive, your subsequent panel concludes with two antibodies that you cannot rule out; lets say Anti-E and Anti-Fya.

Your next step would be to review the histograms of other panels that are in your blood bank (even outdated panels) and look for cells that are positive for Ag-E and negative for Ag-Fya; and look for other cells that are positive for Ag-Fya and negative for Ag-E. Now, here's a question for you; how many positve cells do we need in order to rule in an antibody? It is this number of positive cells you want to accumulate for each of your suspected antibodies individually; ie a cell that is positive for Ag-E and negative for AG-Fya, and vise versa. Please let me know your answer.

[L106]

Trisram,

I think that what Malcolm and others were trying to explain is that after the first panel is complete and you can not rule out two antibodies you must then continue with selected panel cells of panels in your blood bank other then the panel just used. These selected panel cells are cells that you select that are negative for the corresponding antigen of one of your two suspected antibodies and positive for the other; and vise versa. Here's an example, your ABSC is positive, your subsequent panel concludes with two antibodies that you cannot rule out; lets say Anti-E and Anti-Fya.

Your next step would be to review the histograms of other panels that are in your blood bank (even outdated panels) and look for cells that are positive for Ag-E and negative for Ag-Fya; and look for other cells that are positive for Ag-Fya and negative for Ag-E.

Very nice explanation, rravkin.

[rravkin@aol.com]

Very nice explanation, rravkin.

Thank you L106.:):):):):):) or can I call you Donna?

[L106]

Thank you L106.:):):):):):) or can I call you Donna?

You are welcome. You may call me whatever you prefer.