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Antigen Typing QC failed


seraph44
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Hello Blood Bankers,

I recently ran into my first unsatisfactory Antigen typing QC. A tech performed QC for an "e" antisera using a heterozygous cell. Results were negative at IS and negative after 5 min incubation. This was repeated by the same tech and later repeated by myself:Results did not change.

The "e" antisera was ran with a different heterozygous cell and results were negative at IS and 2+ after 5 min incubation. Both the cells and the antisera were near expiration date. However, I'm a bit confused as to what corrective action is needed. How do I prove my panel cells are working correctly and how do I prove my antisera is working correctly?

I think I proved my antisera is working correctly by running it with another Hetero cell. Should I run my panel cell with a new antisera to prove the panel is "e" positive?

Thank you,

Serafin 

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For what it is worth. Years ago we had a panel cell that did not react with commercial anti-e. We were using it as a control for paternity testing. I sent it to one of our suppliers (not the one from whom we purchased the panel). The cell was labeled incorrectly. it was an Rh type that would not normally be on a panel cell.

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I am using ortho for my antisera and ortho for my panel cells. 

I will be testing the cells with a 10 min RT incubation. If that detects it, i'll revise my SOP to 10 minutes like Anorris mentioned. I guess I can notify the ortho about it  as well. 

Thank you,

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On ‎6‎/‎26‎/‎2016 at 11:14 AM, seraph44 said:

Hello Blood Bankers,

I recently ran into my first unsatisfactory Antigen typing QC. A tech performed QC for an "e" antisera using a heterozygous cell. Results were negative at IS and negative after 5 min incubation. This was repeated by the same tech and later repeated by myself:Results did not change.

The "e" antisera was ran with a different heterozygous cell and results were negative at IS and 2+ after 5 min incubation. Both the cells and the antisera were near expiration date. However, I'm a bit confused as to what corrective action is needed. How do I prove my panel cells are working correctly and how do I prove my antisera is working correctly?

I think I proved my antisera is working correctly by running it with another Hetero cell. Should I run my panel cell with a new antisera to prove the panel is "e" positive?

Thank you,

Serafin 

Sometimes antisera does not react the same as a patient sample.   Do you have a patient example of a >e you can run against the cell in question?  I would review all patient reactions with that panel cell and make sure that the cell was not used to rule out anti-e.   I would report this to the manufacturer as well.  I would document all of this.  

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14 hours ago, seraph44 said:

I am using ortho for my antisera and ortho for my panel cells. 

I will be testing the cells with a 10 min RT incubation. If that detects it, i'll revise my SOP to 10 minutes like Anorris mentioned. I guess I can notify the ortho about it  as well. 

Thank you,

With Ortho BioClone Rh antisera, the manufacturer's instructions, require all negative results to have a follow up incubation of 5-10 minutes. I would give it the 10 minutes before I did anything else. We were in the same boat with our anti-e sera, I think they expired yesterday.

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Something similar happened to us about 2 or 3 years ago. One of my techs used a panel cell for the positive C control for antigen typing. The positive control was negative, so she thought she had selected the wrong bottle by mistake. She repeated testing again. Same answer of negative. So she tested the panel cell with anti-C and it was negative. She tested it with a different lot number from the same company and it was also negative. She tested the antisera with a different panel cell to see if it was a problem with the antisera. It wasn't. The panel cell that said it was C+ was not.

So I looked thru all the patient antibody ID to see if we had ruled out Anti-C based on that one panel cell. All was OK. I wrote it all up and filed it away. Emailed Immucor and never heard anything from them.

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First of all, I would read the package insert very carefully.  I do not know this particular reagent, but I do know that sometimes panel cells, depending on what they are suspended in and their concentration, just are not suitable for use as controls with some reagents.  Secondly, a Ee cell had been chosen to test the antiserum.  Some of the posts above were worried that this cell might have been used to rule out an anti-e.  Well, I hope nobody is using Ee cells to rule out the presence of anti-e with ee cells being in such abundance.  Thirdly, you have probably got a second lot of panel cells.  You should test the anti-e with the Ee cell on the other panel.  If that is reacting strongly, then it is possible that the original Ee cell was in fact a previously undetected RH:CE variant and the manufacturer should be notified.  That should not stop you using the cell, however, as you would look at the panel results as a whole, and not just this cell in isolation, and the other antigens (outside the Rh system) would not be affected.  On the other hand, if none of your Ee cells are reacting and you are following the instructions in the package insert to the letter, then you probably do have a problem with you anti-e.  You should stop using it and contact the manufacturer.  They may well need examples of the cells that you are finding negative to see if they can reproduce your reactions on their retained samples. 

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I reported the issue to Ortho and they wanted to point the finger at us vs. the reagent. I concluded it was a combination of both the antisera and the panel cell being near expiration. Even though I used another panel cell that did react at 5 min, it is my secondary panel which is better preserved. 

My corrective action will be to recommend a RT incubation of 10 minutes. This is because I had 2 different hetero cells, which should have reacted at 5 min RT incubation, but only 1 did. What can prove the same thing cannot happen to a short dated unit of blood? I would hate to call it negative after 5 minutes when it really is positive.

Thank you all for your feedback.

Edited by seraph44
grammar
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