We are not getting any reactivity with our Complement Control Cells. They don't expire for another 11 days. We tried 2 different lot numbers of anti-sera and got the same results. We had this same problem about a year ago. Both the anti-C3b, C3d and the check cells are the same brand. Is anyone else experiencing this? It will probably take the maufacturer a year to "recall" them.

I have noticed that when performing anti-Compl testing using tubes, the Immucor control cells react notoriously weak; even after 5 minute incubation I would be lucky to get a really good 1+ reaction. Now I do this testing using Ortho's buffered gel card. The Immucor cells are always 3-4+ with this test system (when used neat or as 0.8%).

Would you share your procedure please?

We noticed that when using the Immucor complement cells with the Ortho anti-C3d, the results were non-existent even after 5-10 minutes incubation at room temperature. At first I thought the problem was with the complement cells, but after ordering Immucor anti-C3d, we noticed the reactions were stronger. After evaluating the Ortho and Immucor anti-C3d we decided to use the Immucor product. I also noticed that the Ortho product was anti-C3d, while the Immucor antisera was a combination of anti-C3d,C3b. Ortho offers an anti-C3d,C3b product, but I never evaluated this product. I figured; why not use Immucor antisera and controlling it with Immucor C3d control

The package insert for the Immucor complement control cells states that "agglutination by anti-complement is seldom as strong as that customarily seen with anti-IgG and is more readily dispersed". Our expected QC result for anti-complement QC is W+-1+. I like this control because it also QCs the techs ability to not "over-shake" in tube testing, and ability to detect weakly reactive agglutination.

Complement Control Cells can also be adversely affected by shipping conditions. Once every couple of years I have a lot # that does not work because they came in when the temperatures were 100 F + . Reactions are extremely weak to non-existent. In the winter time, they have come in too cold, showing traces of hemolysis. Those cells need replaced as well.

I agree with the previous poster. The maximum agglutination you should expect is 2+ with 1+ reactions quite normal. The agglutination is fragile. I, too, find them to be a good test of the shaking abilitiy of my techs and students.

We store our anti-C3d,C3b and complement control cells in the refrigerator and only have them on the bench when in use. I have also split my standing order to get shipments every 2 weeks.

Sue makes an excellent point with refrigeration when not in use.

We only take out the cells when required. The package insert states the complement reactivity deteriorates during the life expectancy of the product.

Mary - I have validated my complement testing using Ortho anti-C3b,-C3d. 50 uL of 0.8% cell suspensions of patient cells and Complement Control cells - using the buffered gel card. 25 uL of the anti-C rgnt to both pt cells and Control cells. I also run a neg control of pt cells and diluent - just to be certain I am not seeing something non-specific to complement activity. Incubate for 5 minutes at room temp. 10 minute spin. Someone mentioned to me in a previous thread that they found the gel did not detect weak reacting anti-C activity. My feelings are that if it is that weak, I would not detect it using classic tube technique anyway, so . . . Hope this is helpful.

We also were seeing non-reactive complement check cells. We began only pulling the cells out at time of need and cleared the problem up. We use combo Ortho and dont seem to have a problem now.

We implemented 5 min RT incubation and it worked fine with the Anti-C3b,-C3d reagent.

Traci

Ah, the old shakedown problem. I visited my BB mentor one day. He was working on a transfusion reaction workup. I asked him if it was the usual febrile reaction. He said no, it was a real antibody. It seems that the patient had a 1+ reactivity, but the tech that did the type and screen had a 2+ shakedown. No joke.

BC