I am working on a procedure for gel antigen typing with anti-kell antisera.

 

It is an antisera where you add AHG... so then is it done in IgG cards?
and the instructions for use says to incubate at 37 for 15 min so that would be followed in the cards as well, but I'm confused to the need of washing like you do with tube... obviously this cannot be done but will it cause an issue by obmiting this process?

 

And since we are going away from the instructions for use, am I correct in saying a validation must be done? Well what if we do no tube testing where I work so I do not have means to compare these two methods, could I instead just test the gel method with heterozygous and negative pannel cells and document this as my validation?

 

Any guidence would be appreciated.

 

Also since this is IgG cards do we suspend the patien/donor red cells in Diluent 2 instead of Diluent 2 Plus?

usually for us when we receive the cards we do validation on each lot number by testing with known positive cells for K Antigen and if it pass it will be ready to use.

 

we will do suspension of red cells with diluent and add in the gel card, incubate and centrifuge then read the result.

Do you have any neighboring reference labs or hospitals that you could share samples and results with for your validation?

It doesn't take much to do that validation.  If you are using a Kell ab that requires ahg testing you will use the IgG card - 15 min at 37 and a 10 minute spin. There is no wash step necessary (you couldn't in the cards anyway).  I use a monoclonal anti-K so I use the buffered gel card with a RT incubation and then the 10 minute spin.  You just need to decide how many samples you want so you feel your validation is valid.  I use Hemobiosciene's diluent which is equivalent to the Diluent 2+.  I try to run homozygous, heterozygous, and negative cells.

Thanks guys! I appreciate the advice.

 

Goodchild... no were the only ones in the area that is going to do antigen typing in gel everyone else does tube.